Methods to identify arthropod-borne pathogens, arthropods, and their blood meal hosts and uses thereof

Abstract

This disclosure is directed to methods to identify arthropod-borne pathogens, arthropod species, and arthropod blood meal hosts. The arthropod-borne pathogen may be a bacterium, a protozoan, or a virus. The arthropod may be a tick which may be a hard or a soft tick. From a single sample, the method uses a combination of PCR and mass spectrometry to rapidly identify a wide variety of pathogens, arthropods, and blood meal hosts. Also provided are methods of diagnosing arthropod-associated diseases in humans, domestic animals, farm animals, or wild animals. Kits are also provided. Furthermore, the methods disclosed herein are useful for a variety of challenges such as arthropod-borne disease surveillance, sentinel surveillance, and monitoring eradication programs.

Description

Methods to identify arthropod-borne pathogens, arthropods, and their blood meal hosts and uses thereofCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U. S. Provisional Appn. No. 63 / 717,098 filed November 6, 2024, Gunselman et al., entitled “Method to identify arthropod-borne pathogens, arthropods, and their blood meal hosts and uses thereof’, Atty. Dkt. 806-02-PROV, which is hereby incorporated by reference in its entirety.REFERENCE TO A “SEQUENCE LISTING,” APPENDIX SUBMITTED IN AN XML FILE FORMAT

[0002] This application contains an ST.26 sequence listing appendix. It has been submitted electronically via EFS-Web as an XML file entitled “806-02-PROV_SEQl.xml”. The ST.26 sequence listing is 521,687 bytes in size and was created on 04-NOV-2024. It is hereby incorporated by reference in its entirety. Applicants certify that the electronic sequence listing and the sequence listing in the specification are the same.1. FIELD

[0003] This disclosure is directed to methods to identify arthropod-borne pathogens, arthropod species, and arthropod blood meal hosts. The arthropod-borne pathogen may be a bacterium, a fungus, a protozoan, or a virus. The vector may be a Culicoides fly, a flea, a louse, a mosquito, a sandfly, a tick, a triatome bug, or a tsetse fly. The tick may be a hard or a soft tick. From a single sample, the method uses a combination of PCR and mass spectrometry to identify a wide variety of vector-borne pathogens, arthropod vectors, and their blood meal hosts. Also provided are methods of diagnosing arthropod-associated diseases in humans, domestic animals, farm animals, or wild animals. Kits are also provided. Further, the methods disclosed herein are useful for a variety of challenges such as arthropod-borne disease surveillance, sentinel surveillance, and monitoring eradication programs.2. BACKGROUND

[0004] INTRODUCTION

[0005] The “background” description provided herein is for the purpose of generally presenting the context of the disclosure. Work of the presently named inventors, to the extent it is described in this background section, as well as aspects of the description which may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art against the present disclosure.

[0006] INTRODUCTION: TICKS

[0007] Infections from tick-borne pathogens are associated with thousands of human illnesses every year. Improved tick surveillance is key for better public health. Tick identification is challenging because of the markedly different appearance at various stages in their life cycle. In most cases the tick may not be found, the intact tick is unavailable, or the tick identification is nearly infeasible because of the life cycle stage, engorged status, or lack of entomological expertise. Nymphs and engorged ticks are difficult even for experts to identify.

[0008] The number of reported tick-borne diseases has steadily increased in recent decades driven by improved tracking, increased deer and tick populations, changing habitats and urbanization, increased human-tick interactions, global climate change, and increased humidity. Many regions have seen the geographic expansion of tick species along with the associated tick-borne diseases. In North America Amblyomma americinum. the lone star tick, has expanded northward, along with its related pathogens. The Gulf Coast tick, Amblyomma maculatum, has expanded well beyond its traditional range and is an important vector of Rickettsia.

[0009] Ticks carrying pathogens cause human diseases including anaplasmosis, babesiosis, Borrelia mayonii infection, Borrelia miyamotii infection, Bourbon virus infection, Colorado tick fever, ehrlichiosis, Heartland virus infection, Lyme disease, Powassan disease, Rickettsia parkeri or 364D rickettsiosis, Rocky Mountain spotted fever (RMSF), southern tick-associated rash illness (STARI), tick-borne relapsing fever (TBRF), and tularemia. The symptoms associated with many of these diseases are hard to distinguish. For example, most patients present with non-specific symptoms such as fever, headache, malaise, and / or myalgia. In addition, many pathogens are either slow growing or difficult to culture. This makes identification of the specific disease and / or pathogen particularly challenging, particularly for serologic assays which may be falsely negative during the first 4-6 weeks after initial infection (CDC, Clinical Testing and Diagnosis for Lyme Disease, 2024). However, treatment of Lyme disease is most effective in its early stage which occurs 3 to 30 days after initial infection, and patients who are not treated in early disease go on to develop severe symptoms such as immune system activity in eye nerves and heart tissue, arthritis, and inflammatory skin disease (Mayo Clinic, Lyme Disease www(dot)mayoclinic(dot)org(slash)diseases-conditions(slash)lyme-disease(slash)symptoms-causes(slash)syc-20374651 dated Feb. 10, 2023 accessed Oct. 3, 2024). Therefore, there is a need in the art for a quick, reliable, specific, and sensitive method for identifying tick-borne pathogens, ticks, and tick blood meal hosts.

[0010] INTRODUCTION: MOSQUITOES

[0011] According to the CDC, mosquitoes are the world’s deadliest animal and kill more people than any other. They list the following vector-borne diseases as spread by mosquitoes:chikungunya, dengue, Eastern equine encephalitis, Japanese encephalitis, La Crosse encephalitis, malaria, St. Louis encephalitis, West Nile, yellow fever, and zika.

[0012] Malaria is a huge public health problem worldwide. The WHO estimates that in 2022 there were 249 million cases of malaria and 608,000 deaths. Over half of those deaths are children under the age of 5. Malaria is spread by the Anopheles mosquito which transmit parasites of the Plasmodium genus. Of the major malarias effecting human health Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, and Plasmodium knowlesi. The vast majority of the cases and deaths are caused by Plasmodium falciparum. Plasmodium vivax is the most widely distributed and was previously considered nontransmissible, but now recognized as capable of spreading malaria. Accurate malaria diagnosis is crucial to reduce presumptive treatments. Currently, microscopy is the main diagnostic tool for malaria together with rapid diagnostic tests (RDT). Current RDTs are lateral flow immunoassays that require a drop of blood as a sample. The RDTs require at least 10 parasites per ul of blood for an accurate diagnosis. They have limited ability to identify asymptomatic carriers who present with low parasitemia. The RDTs are unable to test parasite reservoirs (mosquitos) which is crucial to national malaria elimination programs.

[0013] Another vector-borne pathogen is West Nile virus (WNV), which can cause febrile illness (fever) in some people together with other symptoms such as headache, body aches, joint pains, vomiting, diarrhea, or rash. West Nile symptoms overlap with the symptoms of many other tick-borne diseases making accurate differential diagnosis a challenge for current available technologies.

[0014] Many viruses hosted by mosquitoes or ticks can cause encephalitis. Encephalitis is a rare, potentially life-threating condition due to inflammation of active tissues in the brain. The inflammation leads to brain swelling and adverse neurological consequences. It is frequently due to an infection or an autoimmune reaction. Treatment varies depending on the cause, but most patients require hospitalization. Vector-borne pathogens that cause encephalitis include Chikungunya virus (CHIKV), Dengue virus (DENV), Eastern and Western equine encephalitis virus (EEEV and WEEV), Japanese encephalitis virus (JEV), La Cross virus (LACV), Powassan virus (POWV), St. Louis encephalitis virus (SLEV), Tick-borne encephalitis virus (TBEV), West Nile virus (WNV), and Zika virus (ZIKV). If the cause of the encephalitis is known, physicians and patients can make better treatment decisions.

[0015] INTRODUCTION: FLEAS, FLIES, LICE, ETC.

[0016] Biting midges, Culicoides paraensis, along with mosquitoes are vectors for the Oropouche fever virus (OROV). M. S. Contigiani et al. (2017). Bunyaviruses. In Marcondes, C.B. (Ed.), Arthropod Borne Diseases (pp. 144-145). Springer International Publishing. Fleas are the vector for the plague. Specifically, the flea, Xenopsylla cheopis, is a vector for the bacterium Yesinia pestis, that was responsible for multiple pandemics including the Black Death that killed 25% of the people of Europe in the Middle Ages. Leal-Balbino et al. (2017). Plague and Tularemia. In Marcondes, C. B. (Ed.), Arthropod Borne Diseases (pp. 155). Blood sucking lice are known to spread epidemic typhus (caused by Rickettsia prowazekii), louse-borne relapsing fever (caused by Borrelia recurrenlis). or trench fever (caused by Bartonella quinlana). C. B. Marcondes and P. M. Linardi (2017). Sucking and Chewing Lice. In Marcondes, C. B. (Ed.), Arthropod Borne Diseases (pp. 511-512). Sandflies, Phlebotomus sp., and blackflies, spread Trypanosoma evansi and Leishmania. C. B. Marcondes. Arthropod-Borne Diseases and History. In Marcondes, C. B. (Ed.), Arthropod Borne Diseases (pp. 43-44). Triatomine bugs, Hemiptera sp. and Reduviidae sp., spread Trypanosoma cruzi causing Chagas disease. J. C. P. Dias. Chagas Disease (American Trypanosomiasis). In Marcondes, C. B. (Ed.), Arthropod Borne Diseases (pp.245). Tsetse flies, Glossina sp., are vectors for Trypanosoma brucei causing sleeping sickness in people and nagana disease in livestock. D. Steverding. Sleeping Sickness and Nagana Disease Caused by Trypanosoma brucei. In Marcondes, C. B. (Ed.), Arthropod Borne Diseases (pp. 277-278).

[0017] INTRODUCTION: PATENT AND JOURNAL DISCLOSURES

[0018] Beier discloses an enzyme-linked immunosorbent assay (ELISA) for identifying the blood meal host and the malaria sporozoite. Beier et al. 1988 J Med Entomol 25(1) 9-16. They used their ELISA to analyze Anopheles funestus, Anopheles gambiae, and Anopheles stephensi mosquitoes for blood meal hosts from chicken, cow, dog, goat, horse, human, or pig blood meals. They also used their ELISA to analyze mosquitoes for sporozoites, Plasmodium falciparum and Plasmodium vivax. On page 15, they state that their ELISA method also works for sandflies (Phlebotominae).

[0019] Diarra discloses the proteomic identification of ticks in Mali by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Diarra et al. 2017 PLoS Negl Trop Dis 11(7): e0005762. They found a 99.6% correlation between the MS identification and experts’ morphological identification. They dissected the ticks and used the legs for MS analysis. Half of the tick’s body was used for pathogens screening by PCR. See Diarra at page 5. While they screen some separate tick sections for Anaplasmataceae, Borrelia sp., Coxiella burnetii, and Rickettsia sp., on page 20 they report that their “MALDI-TOF MS analysis was not able to differentiate ticks which were infected or not by the bacteria that were screened.”

[0020] Dietrich discloses a semi-multiplex real-time PCR (RT-PCR) assay for a variety of Borrelia species. Dietrich et al. 2021 J Clin Microbiol 59e02981-20. The assay is designed to detect relapsing fever (RF) Borrelia and classifies them into three groups, a Borrelia hermsii group, a Borrelia miyamoti group, and a Borrelia recurrentis group. The assay uses two primers and five probes to detect Borrelia coriaceae. Borrelia hermsii, Borrelia miyamoloi, Borrelia parkeri, Borrelia recurrentis, o Borrelia turicatae. See Dietrich 2021, Fig. 1, Table 1, and 2.

[0021] Gueye discloses a method to identify the host feeding preferences of malaria vectors in villages in Senegal. Gueye et al. 2023 Sci Reports 13 16410. The mosquitoes were identified morphologically and tested by ELISA using the method of Beier et al. 1988 to identify a chicken, cow, horse, human, or pig blood meal. They identified Anopheles arabiensis, Anopheles coluzzii, Anopheles coustani, Anopheles funestus, Anopheles gambiae, and Anopheles rufipes, mosquitoes. In their study, most mosquitoes had a single species blood meal and a few had multiple species blood meals.

[0022] Modarelli discloses the “TickPath Layerplex” a multiplexed PCR assay for tick-borne diseases (TBDs) in domestic dogs. Modarelli et al., 2019 Sci Reports 9:6950. Briefly, Layerplex is a two-step PCR. They have primers and probes for 11 different pathogens Anaplasma phagocytophilum (Ap); Babesia gibsoni, Babesia canis vogeli, Babesia conradae Ipan-Babesia) Borrelia hermsii (Bh), Borrelia turicatae (Bt), Borrelia parkeri (Bp), Borrelia burgdorferi (Bb); Ehrlichia canis (Ec), Ehrlichia chaffeensis (Ech), Ehrlichia ewingii (Ee), and Rickettsia rickettsii (Rr). Modarelli 2019, page 1, Figure 1. The probes are linked to four different fluorophores which are grouped by pathogen. For example, all of the Borrelia species probes had the same fluorophore, 6-carboxyfluorescein (FAM). If the sample is negative for FAM, then no species of Borrelia is present. If a sample run is positive for FAM, then a Borrelia species is present. Next, the method requires a subsequent followup PCR assay (either single plex or multiplex) be run to identify which particular species of Borrelia is present in the positive sample, e.g., Borrelia burgdorferi (Bb).

[0023] U. S. Patent No. 10,928,384 (Kage) discloses a lateral flow immunoassay for the rapid identification of the microorganism that causes Lyme disease, Borrelia burgdorferi. The assay, named the QuickLyme test, detects the VisE protein of Borrelia burgdorferi.

[0024] WO2014 / 197607 (Chiu et a / .)(“the ‘607 application”) discloses the TickChip™ a microarray assay for identifying tick-borne disease-inducing microorganisms. The TickChip™ is a 57,954 probe assay with 70mer probes to conserved and variable regions of 16S ribosomal RNA for tick-borne bacteria and 18S ribosomal RNA for tick-borne fungi, or protozoans. See the ‘607 application, paragraphs

[0007] ,

[0252] , and

[0264] , Nucleic acids were extracted from wholeblood; amplified by RT-PCR; fluorescent labeled; and hybridized to the microarray. See paragraph

[0266] ,

[0025] The current CDC standards for Lyme disease diagnostic are (i) the standard two-tiered testing (STTT) algorithm and (ii) the modified two-tiered testing (MTTT) algorithm. Both diagnostics are based on antibodies against Borrelia antigens. The STTT uses an enzyme immunoassay (EIA) or chemiluminescence immunoassay (CIA) followed by a western immunoblot. The MTTT uses a first EIA test followed by a second EIA test. See Mead et al., 2019, MMWR 68(32) 703; Madison-Antenucci et al., 2020 Clin Microbiol Rev 33(2) 1-35, 11. Both CDC recommended diagnostic assays have significant shortcomings in that they are unable to detect early infections or reinfections. Current molecular tests, e.g., RNA or DNA-based tests, are single- or low-plexed real-time PCR for one or a limited number of pathogens. PCR tests may be used for detecting Lyme disease and others as laboratory-developed test (LDT).3. SUMMARY OF THE DISCLOSURE

[0026] This disclosure provides an unexpected, surprising improvement over the existing technology with many applications including surveillance and clinical diagnostics. The methods described herein present a vast improvement over existing technology for environmental surveillance and clinical diagnostics. It provides a means to detect vector-borne pathogens, identify vector species, and detect vector hosts — blood meals with a single assay from a single sample. The methods described herein provide valuable information for surveillance by providing rapid information about the occurrence of various vectors, arthropods, e.g., mosquitoes, ticks, their pathogens and their feed sources-blood meals. The methods disclosed herein provide actionable, evidence-based information on infection risk to patients, clinicians, the public, and policy makers on where and when people are at risk for exposure to arthropod-borne pathogens.

[0027] The assay may be used for: 1. Epidemiological surveillance for public health; 2. Patient diagnostic testing for suspected pathogen; 3. Animal health, some pathogens are readily spread among humans and many animals, especially farm animals and domestic animals.

[0028] In the field of clinical diagnostics, the methods disclosed herein provide an approach to detect many pathogens that cause human vector-borne diseases with one assay from various specimen types including blood, CSF, synovial fluid, lesion fluid from a vector bite, erythema migrans biopsy, and other human tissues or body fluids. The assay can detect a number of pathogens within one test from vectors, e.g., a mosquito or tick, bite-wounds, patients' whole blood, synovial fluid, cerebrospinal fluid (CSF), or other body fluids. This assay is the most comprehensive panel detecting these pathogens and their subspecies. This assay can address both malaria clinical diagnoses and malaria / mosquito environmental monitoring needs. This assayincludes most tick-bome pathogens from ticks and mosquitoes. The assay can detect a wide variety of pathogens at the same time.

[0029] The methods are particularly useful with rare or limited samples. The methods provide results in 8 hours offering clinicians timely actionable information for patient treatment thereby reducing misdiagnosis.

[0030] The present disclosure provides a method of identifying (i) an arthropod-borne pathogen, (ii) an arthropod species, and (iii) a blood meal host from a single biological sample, the method comprising: (a) amplifying nucleic acids in the biological sample with a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three arthropod species, and (iii) PCR primers specific for at least three blood meal hosts; (b) performing single base extension with a set of extension primers on the amplified products to generate a set of extended products; and (c) analyzing the set of extended products so as to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).

[0031] A method for diagnosing an arthropod-related disease which comprises: (a) identifying the arthropod and the arthropod-borne pathogen in the biological sample from a human patient or an animal subject by the methods above; and (b) using the identification of the arthropod and the arthropod-borne pathogen to diagnose the arthropod-related disease.

[0032] In yet another embodiment, the disclosure provides a kit for identification of an arthropod species, an arthropod-borne pathogens, or a blood meal hosts from a single biological sample, wherein the kit comprises: (a) a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three species of arthropods, and (iii) PCR primers specific for at least three host species; (b) a set of extension primers; and (c) appropriate reagents and instructions for a user to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).4. BRIEF DESCRIPTION OF THE FIGURES

[0033] FIG. 1 shows a summary slide of the pathogens detected, arthropod identified, and blood meal sources from a series of 116 experiments.

[0034] FIG. 2 shows pathogen identification mass spectrometry plots for a sample that is negative for Amblyomma americanum (Lone star tick) and sample that is positive for Amblyomma americanum. In this figure and subsequent figures (FIG. 3-FIG. 16, FIG. 22-FIG. 30) the DNA was extracted from field-collected ticks were tested and positive samples shown in these figures.

[0035] FIG. 3 shows tick identification mass spectrometry plots for a sample that is negative for Amblyomma maculatum (Gulf Coast tick) and sample that is positive for Amblyomma maculatum.

[0036] FIG. 4 shows pathogen identification mass spectrometry plots for a sample that is negative for Borrelia burgdorferi and sample that is positive for Borrelia burgdorferi.

[0037] FIG. 5 shows pathogen identification mass spectrometry plots for a sample that is negative for Borrelia miyamotoi and sample that is positive for Borrelia miyamotoi.

[0038] FIG. 6 shows pathogen identification mass spectrometry plots for a sample that is negative for generic Borrelia (Borrelia spf and sample that is positive for Borrelia sp.

[0039] FIG. 7 shows tick identification mass spectrometry plots for a sample that is negative for Dermacentor variabilis (American dog tick) and sample that is positive for Dermacentor variabilis.

[0040] FIG. 8 shows pathogen identification mass spectrometry plots for a sample that is negative for Ehrlichia ewingii and sample that is positive for Ehrlichia ewingii.

[0041] FIG. 9 shows pathogen identification mass spectrometry plots for a sample that is negative for Ehrlichia chafeensis and sample that is positive for Ehrlichia chafeensis.

[0042] FIG. 10 shows pathogen identification mass spectrometry plots for a sample that is negative for generic Ehrlichia (Ehrlichia spf and sample that is positive for Ehrlichia sp.

[0043] FIG. 11 shows tick identification mass spectrometry plots for a sample that is negative for Ixodes pacificus (Western black-legged tick) and sample that is positive for Ixodes pacificus.

[0044] FIG. 12 shows tick identification mass spectrometry plots for a sample that is negative for Ixodes scapularis (deer tick) and sample that is positive for Ixodes scapularis.

[0045] FIG. 13 shows pathogen identification mass spectrometry plots for a sample that is negative for Panola Mountain Ehrlichia (PME) and sample that is positive for PME.

[0046] FIG. 14 shows pathogen identification mass spectrometry plots for a sample that is negative for Rickettsia amblyommatis and sample that is positive for Rickettsia amblyommatis.

[0047] FIG. 15 shows pathogen identification mass spectrometry plots for a sample that is negative for Rickettsia parkeri and sample that is positive for Rickettsia parkeri.

[0048] FIG. 16 shows pathogen identification mass spectrometry plots for a sample that is negative for generic Rickettsia (Rickettsia spf and sample that is positive for Rickettsia sp.

[0049] FIG. 17 shows pathogen identification mass spectrometry plots for a sample that is negative for generic Plasmodium species (Plasmodium spf and sample that is positive for Plasmodium sp.

[0050] FIG. 18 shows pathogen identification mass spectrometry plots for a sample that is negative for Plasmodium falciparum and sample that is positive for Plasmodium falciparum.

[0051] FIG. 19 shows pathogen identification mass spectrometry plots for a sample that is negative for Plasmodium knowlesi and sample that is positive for Plasmodium knowlesi.

[0052] FIG. 20 shows pathogen identification mass spectrometry plots for a sample that is negative for Plasmodium ovale and sample that is positive for Plasmodium ovale.

[0053] FIG. 21 shows pathogen identification mass spectrometry plots for a sample that is negative for Plasmodium vivax and sample that is positive for Plasmodium vivax.

[0054] FIG. 22 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for the domestic cat, Felis catus and sample that is positive for Felis catus.

[0055] FIG. 23 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for pig (Sus domesticus) and sample that is positive for Sus domesticus.

[0056] FIG. 24 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for the white tail deer (Odocoileus virginianus) and sample that is positive for Odocoileus virginianus.

[0057] FIG. 25 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for generic Felids and sample that is positive for Felids.

[0058] FIG. 26 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for human (Homo sapiens) and sample that is positive for Homo sapiens.

[0059] FIG. 27 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for rabbit (Oryctolagus cuniculus) and sample that is positive for Oryctolagus cuniculus.

[0060] FIG. 28 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for squirrel (Sciurus carolinensis) and sample that is positive for Sciurus carolinensis.

[0061] FIG. 29 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for vole (Microtus arvalis) and sample that is positive for Microtus arvalis.

[0062] FIG. 30 shows host (blood meal) identification mass spectrometry plots for a sample that is negative for wild boar (Sus scrofa) and sample that is positive for Sus scrofa.5. DETAILED DESCRIPTION OF THE DISCLOSURE

[0063] IN GENERAL

[0064] The disclosure provides a method of identifying (i) an arthropod-borne pathogen, (ii) an arthropod species, and (iii) a blood meal host from a single biological sample, the method comprising: (a) amplifying nucleic acids in the biological sample with a set of polymerase chainreaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three arthropod species, and (iii) PCR primers specific for at least three blood meal hosts; (b) performing single base extension with a set of extension primers on the amplified products to generate a set of extended products; and (c) analyzing the set of extended products so as to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s). The arthropod-borne pathogen may be a tick-borne pathogen, a mosquito-borne pathogen, a biting midge-borne pathogen, a sandfly-borne pathogen, a tsetse fly-borne pathogen, a flea-borne pathogen, a louse-borne pathogen, or a triatomine bug-borne pathogen. The arthropod may be a tick, a mosquito, a biting midge, a sandfly, a tsetse fly, a flea, a louse, or a triatomine bug.

[0065] The set of PCR primers may comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, at least fifteen arthropod-borne pathogens, at least twenty arthropod-borne pathogens, or at least twenty five arthropod-borne pathogens. The set of PCR primers may comprise (ii) PCR primers specific for at least three, at least five, at least eight, or at least ten species of arthropods. The set of PCR primers may comprise (iii) PCR primers specific for at least three, at least five, at least eight, at least ten, at least twelve, or at least fifteen blood meal hosts.

[0066] The tick-borne pathogen may be a bacterium, a fungus, a protozoan, or a virus. In some embodiments, the arthropod-borne pathogen is a bacterium and the PCR primers specific for at least ten arthropod-borne diseases hybridize with a 16S and / or an 18S ribosomal nucleic acid of the bacterium. The bacterium may be an Anaplasma, a Bartonella, a Borrelia, a Coxiella, an Ehrlichia, a Francisella, a Rickettsia, or a Yersinia. Specifically, a bacterium such as Anaplasma bovis, Anaplasma caudatum, Anaplasma centrale, Anaplasma marginale, Anaplasma mesaenterum, Anaplasma odocoilei, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma platys, Bartonella bacilliformis, Bartonella elizabethae, Bartonella henselae, Bartonella quintana, Bartonella vinsonii, Borrelia afzelii, Borrelia americana, Borrelia anserina, Borrelia bissettiae, Borrelia burgdorferi, Borrelia californiensis, Borrelia carolinensis, Borrelia coriaceae, Borrelia garinii, Borrelia hermsii, Borrelia kurtenbachii, Borrelia lanei, Borrelia lonestari, Borrelia lusitaniae, Borrelia mayonii, Borrelia miyamotoi, Borrelia parkeri, Borrelia theileri, Borrelia turicatae, Borrelia valaisiana, Coxiella burnetii, Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia mineirensis, Ehrlichia muris, Ehrlichia muris eauclairensis, Ehrlichia muris-like agent, Ehrlichia ruminantium, Ehrlichia sp., Francisella tularensis, Panola Mountain Ehrlichia, Rickettsia africae (aeschlimannii ), Rickettsia akari, Rickettsia amblyommatis, Rickettsia australis, Rickettsia bellii, Rickettsia CA6269, Rickettsia conorii, Rickettsia felis, Rickettsia heilongjiangensis, Rickettsia helvitica, Rickettsia honei,Rickettsia japonica, Rickettsia massiliae, Rickettsia monacensis, Rickettsia montanesis, Rickettsia parkeri, Rickettsia philippi (strain 364D), Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia sibirica, Rickettsia slovaca, Rickettsia typhi, or Yersinia pestis.

[0067] The arthropod-borne pathogen may be a protozoan such as a Babesia, a Cytauxzoon, a Leishmania, a Plasmodium, a Theileria, or a Trypanosome. Specifically, the protozoan may be Babesia beliceri, Babesia bigemina, Babesia bovis, Babesia caballi, Babesia canis, Babesia catia, Babesia conradae, Babesia crassa, Babesia divergens, Babesia duncani, Babesia felis, Babesia foliatad, Babesia taylori, Babesia gibsoni, Babesia hongkongensis, Babesia jakimovi, Babesia lengau, Babesia major, Babesia microti, Babesia motasi, Babesia occultans, Babesia orientalis, Babesia ovata, Babesia ovis, Babesia perroncitoi, Babesia presentii, Babesia rossi, Babesia trautmanni, Babesia venatorum, Babesia vogeli, Cytauxzoon banethi, Cytauxzoon europaeus, Cytauxzoon felis. Cytauxzoon manul, Cytauxzoon otrantorum, Leishmania aethiopica, Leishmania amazonensis, Leishmania archibaldi, Leishmania braziliensis, Leishmania colombiensis, Leishmania donovani, Leishmania garnhami, Leishmania guyanensis, Leishmania infantum, Leishmania killicki, Leishmania lainsoni, Leishmania lindenbergi, Leishmania major, Leishmania mexicana, Leishmania naifft, Leishmania panamensis, Leishmania peruviana, Leishmania pifanoi, Leishmania shawi, Leishmania tropica, or Leishmania venezuelensis, Plasmodium falciparum, Plasmodium knowlesi, Plasmodium malariae, Plasmodium ovale curtisi, Plasmodium ovale wallikeri, Plasmodium sp., Plasmodium vivax, Theileria annae, Theileria annulata, Theileria buffeli, Theileria equi, Theileria haneyi, Theileria lestoquardi, Theileria luwenshuni, Theileria mutans, Theileria orientalis, Theileria ovis, Theileria parva, Theileria separata, Theileria sergenti, Theileria taurotragi, Theileria uilenbergi, or Theileria velifera, Trypanosoma brucei, Trypanosoma congolense, Trypanosoma cruzi, Trypanosoma evansi, Trypanosoma equiperdum, Trypanosoma rangeli, Trypanosoma simiae, or Trypanosoma vivax.

[0068] The arthropod-borne pathogen may be a virus. The virus may be African horse sickness virus, African swine fever virus, Aino virus (AINV), Akabane virus (AKAV), Alkhurma hemorrhagic fever virus, Barmah forest virus (BFV), Bebaru virus (BENV), Bhanja virus, Bluetongue virus, Bourbon virus, Bovine ephemeral fever virus, Bunyamwera virus (BUNV), Cache Valley virus (CVV), Chandipura virus (CHPV), Chikungunya virus (CHIKV), Colorado tick fever virus, Crimean Congo hemorrhagic fever virus (CCHFV), Dengue virus (DENV), Eastern equine encephalitis virus (EEEV), Edge Hill virus, Equine encephelitis virus, Everglades virus (EVEV), Getah virus (GETV), Guaroa virus (GROV), Heartland virus, Ibaraki virus, Ilheus virus (ILHV), Jamestown Canyon virus (JCV), Japanese encephalitis virus (JEV), Kairi virus, Kasba virus, Kyasunur Forest disease virus (KFDV), La Crosse virus (LACV), Main drain virus(MDV), Mayaro virus (MAYV), Middelburg virus (MIDV), Murray Valley encephalitis virus (MVEV), Nairobi sheep disease virus, Ndumu virus (NDUV), Omsk hemorrhagic fever virus, O'nyong nyong virus (ONNV), Oropouche virus (OROV), Peruvian horse sickness virus, Powassan virus (POWV), Rift Valley fever virus (RVFV), Ross River virus (RRV), Sandfly fever virus, Schmallenberg virus (SBV), Semliki forest virus (SFV), Shuni virus, Sindbis virus (SINV), Snowshoe hare virus (SSHV), St. Louis encephalitis virus (SLEV), Tembusu virus, Thogoto virus, Tick borne encephalitis virus (TBEV), Tyuleniy virus, UNA virus (UNAV), Usutu virus (USUV), Venezualan equine encephalitis virus (VEEV), Vesicular stomatitis virus (VSV), Wesselsbron virus, West Nile virus (WNE), Western equine encephalitis virus (WEEV), Yellow fever virus (YFV), Yunnan virus, or Zika virus (ZIKV).

[0069] The method may have primers for (i) both bacteria and fungi, (ii) both bacteria and protozoans, (iii) both bacteria and viruses, (iv) both fungi and protozoans, (v) both fungi and viruses, (vi) both protozoans and viruses, (vii) bacteria, fungi and protozoans, (viii) bacteria, protozoans, and viruses, (ix) fungi, protozoans, and viruses, or (x) bacteria, fungi, protozoans, and viruses.

[0070] The PCR primers specific for at least three the PCR primers specific for at least three arthropod species hybridize with a gene from a ribosomal 12S subunit, a ribosomal 16S subunit, a ribosomal 18S subunit, or a cytochrome c oxidase unit 1 of the at least three arthropod species. If the arthropod is a tick, it may be a soft tick or a hard tick. The tick species may be Amblyomma americanum, Amblyomma cajennense, Amblyomma maculatum, Amblyomma sp., Amblyomma testudinarium, Amblyomma variegatum, Argas persicus, Bothriocroton hydrosauri, Dermacentor albipictus, Dermacentor andersoni, Dermacentor nitens, Dermacentor occidentalis, Dermacentor reticulatus, Dermacentor sp., Dermacentor variabilis, Haemaphysalis longicornis, Haemaphysalis sp., Hyalomma marinatum, Hyalomma sp., Hyalomma truncation, Ixodes cookei, Ixodes holocyclus, Ixodes pacificus, Ixodes persulcatus, Ixodes ricinus, Ixodes scapularis, Ixodes sp., Ixodes spinipalpis, Margaropus winthemi, Ornithodoros moubata, Ornithodoros rudis, Ornithodoros savignyi, Ornithodoros turicatae, Otobius megnini, Rhipicephalus annlatus, Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Rhipicephalus sanguineus, or Rhipicephalus sp.

[0071] The arthropod may be a mosquito, such as an Aedes, an Anopheles, a Coquillettidia, a Culex, a Culiseta, a Haemagogus, an Ochlerotatus albifasciatus, or a Psorophora mosquito. More specifically, the mosquito may be an Aedes aegypti, Aedes albopictus, Aedes bahamensis, Aedes fulvus pallens, Aedes infirmatus, Aedes sollicitans, Aedes sp., Aedes tormentor, Aedes taeniorhynchus, Aedes triseriatus, Aedes vexans, Anopheles albimanus, Anopheles arabiensis,Anopheles atroparvus, Anopheles barbirostris, Anopheles christyi, Anopheles claviger, Anopheles coluzzii, Anopheles constant, Anopheles culicifacies, Anopheles darlingi, Anopheles dims, Anopheles epiroticus, Anopheles farauti, Anopheles fimestus, Anopheles gambiae, Anopheles maculatus, Anopheles melas, Anopheles merus, Anopheles minimus, Anopheles nimbus, Anopheles pseudopunctipennis, Anopheles quadriannulatus, Anopheles rufipes, Anopheles sacharovi, Anopheles sinensis, Anopheles sp., Anopheles stephensi, Anopheles walker, Coquillettidia perturbans, Coquillettidia sp., Coqudlettidia venezuelensis, Culex biscaynensis, Culex declarator, Culex erraticus, Culex fatigans, Culex pedroi, Culex pipiens, Culex quinquefasciatus, Culex sp., Culiseta dyari, Culiseta inornata, Culiseta melanora, Culiseta sp., Haemagogus capricornii, Haemagogus equinus, Haemagogus janthinomys, Haemagogus lucifer, Haemagogus sp., Ochlerotatus albifasciatus, Ochlerotatus sp., Oehler otatus triser iatus, Psorophora confmnis, Psorophora ferox, or Psorophora sp. Mosquito.

[0072] The arthropod may be a biting midge, a sandfly, or a tsetse fly from the Culicoides sp., Glossina sp., Lutzomyia sp. or Phlebotomus sp. Families. Specifically, the biting midge may be Culicoides actoni, Culicoides adersi, Culicoides brevitarsis, Culicoides fulvus, Culicoides furans, Culicoides grahamii, Culicoides imicola, Culicoides inornatipennis, Culicoides insignis, Culicoides insinuatus, Culicoides mdnei, Culicoides obsoletus, Culicoides oxystoma, Culicoides paraensis, Culicoides phlebotomus, Culicoides schultzei, Culicoides variipennis, or Culicoides wadai. The Glossina sp. tsetse fly is Glossina austeni, Glossina brevipalpis, Glossina jusca, Glossina fuscipes, Glossina longipalpis, Glossina morsitans, Glossina palpalis, Glossina tabaniformis, Glossina tachinoides, or Glossina vanhoofi. The Lutzomyia sp. or Phlebotomus sp. sandfly may be Lutzomyia amazonensis, Lutzomyia anduzei, Lutzomyia aracuchensis, Lutzomyia ayrozai, Lutzomyia carerrai, Lutzomyia christophei, Lutzomyia complexa, Lutzomyia diabolica, Lutzomyia evansi, Lutzomyia flaviscutellata, Lutzomyia gomezi, Lutzomyia hartmanni, Lutzomyia intermedia, Lutzomyia llanosmartinsi, Lutzomyia longipalpis, Lutzomyia migonei, Lutzomyia panamensis, Lutzomyia paraensis, Lutzomyia peruensis, Lutzomyia pessoai, Lutzomyia trapidoi, Lutzomyia umbratilis, Lutzomyia wellcomei, Lutzomyia whitmani, Lutzomyia whitmani, Lutzomyia ylephiletor, Lutzomyia ylephiletor, Lutzomyia yucumensis, Lutzomyia olmeca, Phlebotomus alexandri, Phlebotomus ansarii, Phlebotomus argentipes, Phlebotomus ariasi, Phlebotomus caucasicus, Phlebotomus celiae, Phlebotomus chinensis, Phlebotomus duboseqi, Phlebotomus kandelakii, Phlebotomus langeroni, Phlebotomus longicuspis, Phlebotomus longiductus, Phlebotomus longipes, Phlebotomus martini, Phlebotomus near rossi, Phlebotomus neglectus, Phlebotomus orientalis, Phlebotomus papatasi, Phlebotomus pedifer, Phlebotomusperfiliewi, Phlebotomus perniciosus, Phlebotomus sale hi, Phlebotomus sergenti, Phlebotomus smirnovi, Phlebotomus tobbi, Phlebotomus transcaucasicus, o Phlebotomus vansomeranae.

[0073] The arthropod may be a flea such as Aetheca wagneri, Amphipsylla sp., Anomiopsyllus sp., Atyphloceras sp., Callopsylla sp., Catallagia sp., Ceratophyllus sp., Chiastopsylla sp., Citellophilus sp., Coptopsylla sp., Craneopsylla sp., Ctenocephalides sp., Ctenophthalmus sp., Diamanus montanus, Dinopsyllus sp., Eumolpianus eumolpi, Foxella sp., Frontopsylla sp., Hectopsylla sp., Hoplopsyllus sp., Hystrichopsylla sp., Listropsylla sp., Malaraeus sp., Megabothris sp., Megarthroglossus sp., Meringis sp., Monopsyllus sp., Neopsylla sp., Neotyphloceras sp., Nosopsyllus sp., Odontopsyllus sp., Opisocroslis spp., Opisodasys sp., Orchopeas sp., Oropsylla sp., Parapsyllus sp., Pleochaetis sp., Polygenis sp., Psocopsylla sp., Pulex irritans, Rhadinopsylla sp., Stivalius sp., Synosternus sp., Thrassis sp., Tiamastus sp., Tritopsylla sp., Xenopsylla cheopis, or Xenopsylla sp. Alternatively, the arthropod may be a louse such as a Haematopinidae, Hoplopleuridae, Linognathidae, Pediculidae, Polyplacidae, or Pthiridae family member. Specifically, the louse may be Haematopinus asini, Haematopinus eurysternus, Haematopinus quadripertusus, Haematopinus suis, Haematopinus tuberculatus, Haematopinus tuberculatus, Haemodipsus ventricosus, Hoplopleura captiosa, Hoplopleura pacifica, Linognathus africanus, Linognathus africanus, Linognathus ovillus, Linognathus pedalis, Linognathus setosus, Linognathus stenopsis, Linognathus vituli, Pediculus humanus capitis, Pediculus humanus humanus, Polyplax serrata, Polyplax spinulosa, Pthirus pubis, Solenopotes capillatus. In some embodiments, the arthropod may be a triatomine bug such as Panstrongylus megislus, Rhodnius prolixus, Triatoma brasiliensis, Triatoma dimidiate, or Triatoma inf e stans.

[0074] In the methods above, the PCR primers specific for at least three blood meal hosts species may hybridize with a satellite DNA, a tandem repeat, or a mitochondrial gene from the blood meal host species. The blood-meal host species may be an ape, a bat, a bear, a bird, a bovine, a camelid, a canid, a cervid, a dog, an equid, a felid, a fox, a giraffe, a goat, a horse, a human, a monkey, an opossum, a pig, a rabbit, a raccoon, a rodent, a sheep, a shrew, or a wild boar. The bird may be a chicken, a crane, a duck, an emu, a goose, a pheasant, a quail, or a turkey. The bovine may be an auroch, a bison, a buffalo, a domestic cow, an eland, a four-horned antelope, a guar, a spiral-homed antelope, a waterbuck, or a yak. The camelid may be an alpaca, a Bactrian camel, a dromedary, a guanaco, a llama, or a vicuna. The canid may be a coyote, a dog, a jackal, or a wolf. The cervid may be a brocket, a chital, an elk, a montjac, a moose, a mule deer, a red deer, a reindeer, a roe deer, a seka deer, or a white-tailed deer. The equid may be a donkey, a horse, a mule, or a zebra. The felid may be a bobcat, a cougar, a domestic cat, a jaguar, a leopard,a lion, a lynx, a mountain lion, an ocelot, a panther, a puma, or a tiger. The rodent may be a capybara, a chipmunk, a gerbil, a marmot, a mouse, a prairie dog, a rat, a squirrel, or a vole. The mouse is a North American deer mouse or a white-footed mouse.

[0075] Preferably, the extended products are analyzed by mass spectrometry, e.g., MALDI-TOF mass spectrometry. For pathogen identification, the PCR primers may be selected from nucleotide sequences set forth in SEQ ID Nos: 2, 3, 10, 11, 12, 13, 15, 16, 20, 21, 23, 24, 25, 26, 28, 29, 33, 34, 40, 41, 42, 44, 46, 47, 53, 54, 57, 60, 61, 62, 63, 69, 70, 74, 75, 81, 82, 84, 85, 86, 89, 90, 91, 92, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 107, 108, 109, 111, 112, 115, 116, 118, 119, 120, 122, 123, 126, 127, 128, 129, 131, 132, 134, 136, 137, 138, 139, 140, 141, 142, 144, 145, 146, 148, 149, 151, 152, 153, 154, 156, 157, 160, 161, 166, 167, 169, 170, 173, 174, 178, 179, 180, 181, 182, 183, 186, 187, 188, 189, 194, 195, 200, 201, 203, 204, 206, 207, 209, 210, 212, 213, 215, 216, 218, 219, 221, 222, 224, 225, 226, 228, 229, 231, 232, 236, 237, 239, 240, 242, 243, 244, 245, 248, 249, 252, 253, 256, 257, 260, 261, 263, 264, 265, 266, 269, 270, 273, 274, 277, 278, 280, 281, 282, 283, 286, 287, 289, 290, 293, 294, 298, 299, 302, 303, 305, 306, 307, 308, 310, 311, 315, 316, 320, 321, 324, 325, 328, 329, 332, 333, 336, 337, 339, 340, 341, 342, 346, 347, 348, 349, 351, 352, 355, 356, 357, 358, 359, 360, 361, 362, 366, 367, 368, 369, 373, 374, 375, or 376. For arthropod identification, the PCR primers may be selected from nucleotide sequences set forth in SEQ ID Nos: 378, 379, 381, 382, 385, 386, 388, 389, 390, 392, 393, 397, 398, 401, 402, 403, 404, 407, 408, 411, 412, 415, 416, 417, or 418. For host (blood meal) identification, the PCR primers may be selected from nucleotide sequences set forth in SEQ ID Nos: 421, 422, 425, 426, 427, 428, 429, 430, 433, 434, 436, 437, 438, 439, 443, 444, 445, 446, 450, 451, 452, 453, 454, 456, 457, 458, 462, 463, 464, 465, 468, 469, 472, 473, 476, 477, 480, 481, 484, 485, 487, 488, 490, 491, 494, 495, 498, 499, 501, 502, 504, 505, 507, 508, 509, 510, 511, 512, 514, 515, 518, 519, 520, 522, 523, 524, 525, 529, 530, 531, 532, 535, 536, 538, 539, 541, 542, 543, 544, 546, 547, 548, 549, 553, 554, 555, 556, 558, 559, 560, 561, 562, 563, 566, 567, 568, or 569.

[0076] For pathogen identification, the extension primers may be selected from nucleotide sequences set forth in SEQ ID NOs: 1, 8, 9, 14, 19, 22, 30, 31, 32, 39, 43, 45, 48, 49, 50, 52, 55, 58, 64, 65, 66, 67, 68, 71, 72, 76, 80, 83, 88, 93, 106, 110, 113, 114, 117, 121, 124, 125, 130, 133, 135, 143, 147, 150, 155, 158, 159, 165, 168, 172, 175, 177, 184, 185, 192, 193, 199, 202, 205, 208, 211, 214, 217, 220, 223, 227, 230, 235, 238, 241, 246, 247, 250, 251, 254, 259, 262, 268, 272, 276, 279, 285, 288, 292, 296, 297, 301, 304, 309, 314, 317, 319, 322, 323, 330, 331, 335, 338, 343, 344, 345, 350, 353, 354, 363, 364, 365, 370, 371, 372, 377. For arthropod identification, the extension primers may be selected from nucleotide sequences set forth in SEQ ID NOs: 380,383, 391, 394, 396, 399, 405, 409, or 413. For host (blood meal) identification, the extension primers may be selected from nucleotide sequences set forth in SEQ ID NOs: 419, 423, 424, 431, 432, 435, 440, 441, 442, 447, 449, 455, 459, 461, 466, 471, 475, 479, 483, 486, 489, 493, 496, 497, 500, 503, 506, 513, 517, 521, 526, 527, 534, 537, 540, 545, 550, 552, 557, 564, or 565.

[0077] Alternatively, the amplified products or the extended products may be analyzed by next generation sequencing. The amplified products for pathogen identification may be selected from nucleotide sequences set forth in SEQ ID NOs: 4, 5, 6, 7, 17, 18, 27, 35, 36, 37, 38, 51, 56, 59, 73, 77, 78, 79, 87, 104, 105, 162, 163, 164, 171, 176, 190, 191, 196, 197, 198, 233, 234, 255, 258, 267, 271, 275, 284, 291, 295, 300, 312, 313, 318, 326, 327, or 334. The amplified products for arthropod identification may be selected from nucleotide sequences set forth in SEQ ID NOs: 384, 387, 395, 400, 406, 410, or 414. The amplified products for host (blood meal) identification may be selected from nucleotide sequences set forth in SEQ ID NOs: 420, 448, 460, 467, 470, 474, 478, 482, 492, 516, 528, 533, or 551.

[0078] Other embodiments provide an oligonucleotide comprising or consisting of a sequence of nucleotides selected from SEQ ID NOs: 1-566, or a complement thereof. In another embodiment, the present disclosure provides an oligonucleotide that includes a nucleic acid having at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, 99.5% etc.) to one of SEQ ID NOs: 1-478, or a complement thereof. Generally, these oligonucleotides may be primer nucleic acids, probe nucleic acids, extension primer nucleic acids, amplified products (or amplicons) or the like in these embodiments. In certain of these embodiments, the oligonucleotides have 40 or fewer nucleotides {e.g., 35 or fewer nucleotides, 30 or fewer nucleotides, 25 or fewer nucleotides, 20 or fewer nucleotides, 15 or fewer nucleotides, etc.). For the sequences in which the amplicon, extension primer, and PCR forward and reverse primers disclosed herein, the disclosure includes primers are extended by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in the 3’ end or primers extended by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in the 5’ end. Alternatively, this disclosure includes primers that primers are shortened by 1, 2, 3, 4, or 5 nucleotides in the 3’ end or primers shortened by 1, 2, 3, 4, or 5 nucleotides in the 5’ end. The invention also includes pluralities of nucleotides, e.g. any 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 175, 200, 225, 250, 275, 300 or 350 nucleotides.

[0079] In some embodiments, the oligonucleotides comprise at least one modified nucleotide, e.g., to alter nucleic acid hybridization stability relative to unmodified nucleotides. In some embodiments, the oligonucleotides include at least one conservatively modified variation

[0080] The biological sample may be a biopsy, a blood sample, a cerebral spinal fluid (CSF) sample, an environmental sample, a fluid from a skin blister or ooze from a lesion, a milk sample, a saliva sample, a sewage sample, a synovial fluid sample, a tick homogenate, a tissue sample, or a urine sample. The blood sample may be a plasma sample, a peripheral blood mononuclear cell (PBMC) sample, a serum sample, or a whole blood sample.

[0081] In some embodiments, the method is able to identify an arthropod-borne pathogen in a 1 mL blood sample if greater than 10 arthropod-borne pathogen genomes are present in the blood sample. In other embodiments, multiple arthropod-borne pathogens may be present in the biological sample. The biological sample may be a pooled sample or a sample from a single subject or a single arthropod.

[0082] The method provides identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 24 hours. More preferably, the method provides identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 12 hours or less than 8 hours.

[0083] The disclosure also provides a method for diagnosing a vector-borne disease, such as an arthropod-related disease which comprises: (a) identifying the arthropod and the arthropod-borne pathogen in the biological sample from a human or animal patient by the methods disclosed herein; and (b) using the identification of the arthropod and the arthropod-borne pathogen to diagnose the arthropod-related disease. The arthropod-related disease may be a tick-related disease such as anaplasmosis, babesiosis, ehrlichiosis, Lyme disease, rickettsiosis, Rocky Mountain Spotted Fever, tick-borne relapsing fever, or tularemia. The arthropod-related disease may be a mosquito-borne disease. The arthropod-borne disease may be African swine fever, Alkhurma hemorrhagic fever, anaplasmosis, babesiosis, bluetongue disease, Chagas disease, chikungunya, dengue, ehrlichiosis, encephalitis, equine encephalitis, hepatozoonosis, Lyme disease, malaria, Oropouche fever, plague, rickettsiosis, Rocky Mountain Spotted Fever, tick-borne relapsing fever, tularemia, West Nile, yellow fever, or zika fever. The human patient (i) may have experienced an arthropod bite; (ii) may be suspected of having been bitten by an arthropod; or (iii) may be experiencing fever / chills, joint pain, muscle aches, persistent fever, or rash. Arthropod-borne diseases affect many animals including domestic animals, such as dogs or cats; farm animals such as chickens, cows, goats, horses, pigs; or wild animals.. For example, Lyme disease symptoms in dogs include lethargy, lameness, fever, joint pain or swelling, and swollen lymph nodes. Common tick-borne animal diseases include anaplasmosis, babesiosis, ehrlichiosis, or hepatozoonosis.

[0084] The arthropod-borne disease may be an acute or a chronic tick-borne disease.

[0085] A method of disease vector surveillance is also provided which comprises analyzing biological samples by the methods disclosed herein order to identify arthropod vector species or an animal host that may participate in the pathogen life cycle. The disclosure provides a method of sentinel surveillance which comprises analyzing biological samples from a sentinel source by the methods herein in order to identify the arthropod species, the arthropod-borne pathogen(s), and / or the blood meal hosts in the sentinel source. Moreover, the disclosure provides methods of monitoring an eradication program which comprises analyzing biological samples by the methods herein in order to identify the arthropod species, the arthropod-borne pathogen(s), and the blood meal hosts.

[0086] Lastly, the disclosure provides a kit for identification of arthropod species, arthropod-borne pathogens, or a blood meal hosts from a single biological sample, wherein the kit comprises: (a) a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least three species of arthropods, (ii) PCR primers specific for at least ten arthropod-borne pathogens, and (iii) PCR primers specific for at least three host species; (b) a set of extension primers; and (c) appropriate reagents and instructions for a user to identify the arthropod species, the arthropod-borne pathogen, and the blood meal host.

[0087] DEFINITIONS

[0088] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0089] Throughout the present specification, the terms “about” and / or “approximately” may be used in conjunction with numerical values and / or ranges. The term “about” is understood to mean those values near to a recited value. For example, “about 40 [units]” may mean within ± 25% of 40 (e.g., from 30 to 50), within ± 20%, ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, less than ± 1%, or any other value or range of values therein or there below. Alternatively, depending on the context, the term “about” may mean ± one half a standard deviation, ± one standard deviation, or ± two standard deviations. Furthermore, the phrases “less than about [a value]” or “greater than about [a value]” should be understood in view of the definition of the term “about” provided herein. The terms “about” and “approximately” may be used interchangeably. In some embodiments, the term "about" as used refers to a numerical value plus or minus 10% of the numerical value.

[0090] Throughout the present specification, numerical ranges are provided for certain quantities. It is to be understood that these ranges comprise all subranges therein. Thus, the range“from 50 to 80” includes all possible ranges therein e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70, etc.). Furthermore, all values within a given range may be an endpoint for the range encompassed thereby (e.g., the range 50-80 includes the ranges with endpoints such as 55-80, SO-75, etc.).

[0091] As used herein, the term "amplifying" refers to the process of synthesizing nucleic acid molecules that are complementary to one or both strands of a template nucleic acid molecule {e.g., nucleic acid molecules from the pathogen, tick, or blood meal host genome). Amplifying a nucleic acid molecule typically includes denaturing the template nucleic acid, annealing primers to the template nucleic acid at a temperature that is below the melting temperatures of the primers, and enzymatically elongating from the primers to generate an amplification product. Amplification typically requires the presence of deoxyribonucleoside triphosphates, a DNA polymerase enzyme (e.g., Platinum® Taq) and an appropriate buffer and / or co-factors for optimal activity of the polymerase enzyme {e.g., MgCh and / or KC1).

[0092] Throughout the specification the word “comprising,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. The present disclosure may suitably “comprise”, “consist of’, or “consist essentially of’, the steps, elements, and / or reagents described in the claims.

[0093] In some embodiments, the oligonucleotides include at least one conservatively modified variation. "Conservatively modified variations" or, simply, "conservative variations" of a particular nucleic acid sequence refers to those nucleic acids, which encode identical or essentially identical amino acid sequences, or, where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. One of skill in the art will recognize that individual substitutions, deletions or additions which alter, add or delete a single nucleotide or a small percentage of nucleotides (typically less than 5%, more typically less than 4%, 2% or 1%) in an encoded sequence are "conservatively modified variations" where the alterations result in the deletion of an amino acid, addition of an amino acid, or substitution of an amino acid with a chemically similar amino acid.

[0094] As used herein, "elevation" of a measured level of a biomarker relative to a standard level means that the amount or concentration of a biomarker in a sample is sufficiently greater in a subject relative to the standard to be detected by the methods described herein. For example, elevation of the measured level relative to a standard level may be any statistically significant elevation which is detectable. Such an elevation may include, but is not limited to, about a 1%,about a 10%, about a 20%, about a 40%, about an 80%, about a 2-fold, about a 4-fold, about an 8-fold, about a 20-fold, or about a 100-fold elevation, or more, relative to the standard.

[0095] The term "extension" or "elongation" when used with respect to nucleic acids refers to when additional nucleotides (or other analogous molecules) are incorporated into the nucleic acids, n extension reaction is conducted under extension conditions, and a variety of such conditions are known and selected for a particular application. Extension conditions can include certain reagents, including without limitation, one or more oligonucleotides, extension nucleotides (e.g., nucleotide triphosphates (dNTPs)), chain terminating reagents or nucleotides (e.g., one or more dideoxynucleotide triphosphates (ddNTPs) or acyclic terminators), one or more salts (e.g., magnesium-containing salt), one or more buffers (e.g., with beta-NAD, Triton X-100), and one or more polymerizing agents (e.g., DNA polymerase, RNA polymerase). Extension can be conducted under isothermal conditions or under non-isothermal conditions (e.g., thermocycled conditions), in certain embodiments. One or more nucleic acid species can be extended in an extension reaction and one or more molecules of each nucleic acid species can be extended. A nucleic acid can be extended by one or more nucleotides.

[0096] Primer extension processes include methods such as iPLEX™ or homogeneous MassExtend® (hME) (see, for example, U. S. Published Patent Application No. 2013 / 0237428 Al, U. S. Patent No. 8,349,566, and U. S. Patent No. 8,003,317, the contents of which are incorporated in their entirety by reference herein), in which a mixture of minor nucleic acid species (e.g., mutant alleles) and major nucleic acid species (e.g., wild type alleles) are subjected to a polymerase chain reaction (PCR) amplification using a set of amplification primers, a polymerase and deoxynucleotides (dNTPs), thereby generating amplicons of the wild type and mutant species. After treatment with shrimp alkaline phosphatase (SAP) to dephosphorylate unincorporated dNTPs, the amplicon mixture is extended using extension primers (unextended primers or UEPs), a polymerase and a termination mix that includes chain terminating reagents (e.g., dideoxunucleotides or ddNTPs). The UEPs hybridize to the amplicons and are extended either up to the site of variance between the mutant and wild type species (i.e., extension stops at the mutation site where there is a difference in bases between the mutant and wild type species to generate single base extension products or SBEs, as in iPLEX™) or a few bases (e.g., 2-3 bases) past the site of variance (as in, for example, the hME method). The resulting extension products can then be processed {e.g., by desalting prior to mass spectrometry) and analyzed for the presence of the mutant alleles based on a difference in detection signal (e.g., mass) relative to the wild type allele. The above-described iPLEX™ and homogeneous MassExtend® (hME) methods use anequimolar mixture of ddNTPs in the extension step for generating extension products corresponding to wild type and mutant species.

[0097] The term "hybridizing" refers to the annealing of one or more probes to an amplification product. "Hybridization conditions" typically include a temperature that is below the melting temperature of the probes but that avoids non-specific hybridization of the probes.

[0098] The terms "identical" or percent "identity" in the context of two or more nucleic acid sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides that are the same, when compared and aligned for maximum correspondence, e.g., as measured using one of the sequence comparison algorithms available to persons of skill or by visual inspection. Exemplary algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST programs, which are described in, e.g., Altschul et al. (1990) "Basic local alignment search tool" J Mol Biol 215:403-410; Gish etal. (1993) "Identification of protein coding regions by database similarity search" Nature Genet.3:266-272; Madden et al. (1996) "Applications of network BLAST server" Meth Enzymol 266: 131-141; Altschul et al. (1997) "Gapped BLAST and PSLBLAST: a new generation of protein database search programs" Nucleic Acids Res 25:3389-3402; and Zhang et al. (1997) "PowerBLAST: A new network BLAST application for interactive or automated sequence analysis and annotation" Genome Res 7:649-656.

[0099] A "modified nucleotide" in the context of an oligonucleotide refers to an alteration in which at least one nucleotide of the oligonucleotide sequence is replaced by a different nucleotide that provides a desired property to the oligonucleotide. Exemplary modified nucleotides that can be substituted in the oligonucleotides described herein include, e.g., a t-butyl benzyl, a C5-methyl-dC, a C5-ethyl-dC, a C5-methyl-dU, a C5-ethyl-dU, a 2,6-diaminopurine, a C5- propynyl-dC, a C5-propynyl-dU, a C7-propynyl-dA, a C7-propynyl-dG, a C5- propargylamino-dC, a C5-propargylamino-dU, a C7-propargylamino-dA, a C7- propargylamino-dG, a 7-deaza-2-deoxyxanthosine, a pyrazolopyrimidine analog, a pseudo- dU, a nitro pyrrole, a nitro indole, 2'-0-methyl ribo-U, 2'-0-methyl ribo-C, an N4-ethyl-dC, an N6-methyl-dA, a 5-propynyl dU, a 5-propynyl dC, 7-deaza-deoxy guanosine (deaza G (u-deaza)) and the like. Many other modified nucleotides that can be substituted in the oligonucleotides are referred to herein or are otherwise known in the art. In certain embodiments, modified nucleotide substitutions modify melting temperatures (Tm) of the oligonucleotides relative to the melting temperatures of corresponding unmodified oligonucleotides. To further illustrate, certain modified nucleotide substitutions can reduce non-specific nucleic acid amplification (e.g., minimize primer dimer formation or the like), increase the yield of an intended target amplicon, and / or the like in some embodiments. Examplesof these types of nucleic acid modifications are described in, e.g., U. S. Patent No. 6,001,611. Other modified nucleotide substitutions may alter the stability of the oligonucleotide, or provide other desirable features.

[0100] The term "primer" as used herein is known to those skilled in the art and refers to oligomeric compounds, primarily to oligonucleotides but also to modified oligonucleotides that are able to "prime" DNA synthesis by a template-dependent DNA polymerase, i.e., the 3'-end of the, e.g., oligonucleotide provides a free 3' -OH group where further "nucleotides" may be attached by a template-dependent DNA polymerase establishing 3' to 5' phosphodiester linkage whereby deoxynucleoside triphosphates are used and whereby pyrophosphate is released.

[0101] As used herein, the term “reference set” may be an internal, external, or a universal reference set of nucleic acids or expression products used to calibrate a particular sample. For example, an internal reference set of nucleic acids may be obtained using normal tissue or a blood sample from the subject. Alternatively, an internal reference set may based on the total RNA in the sample. In another embodiment, the reference set may be a set of one or more housekeeping genes, e.g., human acidic ribosomal protein (HuPO), P-actin (BA), cyclophylin (CYC), glyceraldehyde-3 -phosphate dehydrogenase (GAPDH), phosphoglycerokinase (PGK), P2-microglobulin (B2M), P-glucuronidase (GUS), hypoxanthine phosphoribosyltransferase (HPRT), transcription factor HD TATA binding protein (TBP), transferrin receptor (TfR), human acidic ribosomal protein (HuPO), elongation factor-l-a (EF-l-a), metastatic lymph node 51(MLN51), or ubiquitin conjugating enzyme (UbcH5B). See Dheda et al. 2004 BioTechniques 37:112-119. An external reference set may be obtained from clinical studies to determine normal ranges and ranges. Alternatively, the reference set may be based on a particular patient population such as smokers, gender, or race. In yet another embodiment, the reference set may be a universal reference set. Many commercial vendors sell cDNA and RNA reference sets of genes or reference libraries.

[0102] The term "thermostable polymerase" refers to a polymerase enzyme that is heat stable, i.e., the enzyme catalyzes the formation of primer extension products complementary to a template and does not irreversibly denature when subjected to the elevated temperatures for the time necessary to effect denaturation of double-stranded template nucleic acids. Generally, the synthesis is initiated at the 3' end of each primer and proceeds in the 5' to 3' direction along the template strand. Thermostable polymerases have been isolated from Thermus flavus, T. ruber, T. thermophilus, T. aquaticus, T. lacteus, T. rubens, Bacillus stearothermophilus, and Methanothermus fervidus. Nonetheless, polymerases that are not thermostable also can be employed in PCR assays provided the enzyme is replenished, if necessary. The term "complementthereof refers to nucleic acid that is both the same length as, and exactly complementary to, a given nucleic acid.

[0103] It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation.

[0104] The term "standard level" as used herein refers to a baseline level of a biomarker as determined in one or more normal subjects. For example, a baseline may be obtained from at least one subject and preferably is obtained from an average of subjects (e.g., n=2 to 100 or more), wherein the subject or subjects have no prior history of disease.

[0105] Any suitable technology can be used to detect and / or quantify amplicons. Non-limiting examples of technologies that can be utilized to detect and / or quantify amplicons include primer extension assays, amplification (e.g., digital PCR, quantitative polymerase chain reaction (qPCR)), sequencing (e.g., nanopore sequencing, massive parallel sequencing), mass spectrometry, array hybridization (e.g., microarray hybridization; gene-chip analysis), flow cytometry, gel electrophoresis (e.g., capillary electrophoresis), cytofluorimetric analysis, fluorescence microscopy, confocal laser scanning microscopy, laser scanning cytometry, affinity chromatography, manual batch mode separation, electric field suspension, the like and combinations of the foregoing.

[0106] Computing Devices

[0107] A computing device may be implemented in programmable hardware devices such as processors, digital signal processors, central processing units, field programmable gate arrays, programmable array logic, programmable logic devices, cloud processing systems, or the like. The computing devices may also be implemented in software for execution by various types of processors. An identified device may include executable code and may, for instance, comprise one or more physical or logical blocks of computer instructions, which may, for instance, be organized as an object, procedure, function, or other construct. Nevertheless, the executable of an identified device need not be physically located together but may comprise disparate instructions stored in different locations which, when joined logically together, comprise the computing device and achieve the stated purpose of the computing device. In another example, a computing device may be a server or other computer located within a hospital or out-patient environment and communicatively connected to other computing devices (e.g., POS equipment or computers) for managing accounting, purchase transactions, and other processes within the hospital or out-patient environment. In another example, a computing device may be a mobile computing device suchas, for example, but not limited to, a smart phone, a cell phone, a pager, a personal digital assistant (PDA), a mobile computer with a smart phone client, or the like. In another example, a computing device may be any type of wearable computer, such as a computer with a head-mounted display (HMD), or a smart watch or some other wearable smart device. Some of the computer sensing may be part of the fabric of the clothes the user is wearing. A computing device can also include any type of conventional computer, for example, a laptop computer or a tablet computer. A typical mobile computing device is a wireless data access-enabled device (e.g., an iPHONE® smart phone, a BLACKBERRY® smart phone, a NEXUS ONE™ smart phone, an iPAD® device, an APPLE WATCH®, smart watch, or the like) that is capable of sending and receiving data in a wireless manner using protocols like the Internet Protocol, or IP, and the wireless application protocol, or WAP. This allows users to access information via wireless devices, such as smart watches, smart phones, mobile phones, pagers, two-way radios, communicators, and the like. Wireless data access is supported by many wireless networks, including, but not limited to, Bluetooth, Near Field Communication, CDPD, CDMA, GSM, PDC, PHS, TDMA, FLEX, ReFLEX, iDEN, TETRA, DECT, DataTAC, Mobitex, EDGE and other 2G, 3G, 4G, 5G, and LTE technologies, and it operates with many handheld device operating systems, such as PalmOS, EPOC, Windows CE, FLEXOS, OS#, JavaOS, iOS# and Android. Typically, these devices use graphical displays and can access the Internet (or other communications network) on so-called mini- or microbrowsers, which are web browsers with small file sizes that can accommodate the reduced memory constraints of wireless networks. In a representative embodiment, the mobile device is a cellular telephone or smart phone or smart watch that operates over GPRS (General Packet Radio Services), which is a data technology for GSM networks or operates over Near Field Communication e.g. Bluetooth. In addition to a conventional voice communication, a given mobile device can communicate with another such device via many different types of message transfer techniques, including Bluetooth, Near Field Communication, SMS (short message service), enhanced SMS (EMS), multi-media message (MMS), email WAP, paging, or other known or later-developed wireless data formats. Although many of the examples provided herein are implemented on smart phones, the examples may similarly be implemented on any suitable computing device, such as a computer.

[0108] An executable code of a computing device may be a single instruction, or many instructions, and may even be distributed over several different code segments, among different applications, and across several memory devices. Similarly, operational data may be identified and illustrated herein within the computing device, and may be embodied in any suitable form and organized within any suitable type of data structure. The operational data may be collected as asingle data set, or may be distributed over different locations including over different storage devices, and may exist, at least partially, as electronic signals on a system or network.

[0109] The described features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. In the following description, numerous specific details are provided, to provide a thorough understanding of embodiments of the disclosed subject matter. One skilled in the relevant art will recognize, however, that the disclosed subject matter can be practiced without one or more of the specific details, or with other methods, components, materials, etc. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosed subject matter.

[0110] As used herein, the term “memory” is generally a storage device of a computing device. Examples include, but are not limited to, read-only memory (ROM) and random access memory (RAM).

[0111] The device or system for performing one or more operations on a memory of a computing device may be a software, hardware, firmware, or combination of these. The device or the system is further intended to include or otherwise cover all software or computer programs capable of performing the various heretofore-disclosed determinations, calculations, or the like for the disclosed purposes. For example, exemplary embodiments are intended to cover all software or computer programs capable of enabling processors to implement the disclosed processes. Exemplary embodiments are also intended to cover any and all currently known, related art or later developed non-transitory recording or storage mediums (such as a CD-ROM, DVD-ROM, hard drive, RAM, ROM, floppy disc, magnetic tape cassette, etc.) that record or store such software or computer programs. Exemplary embodiments are further intended to cover such software, computer programs, systems and / or processes provided through any other currently known, related art, or later developed medium (such as transitory mediums, carrier waves, etc.), usable for implementing the exemplary operations disclosed below.

[0112] In accordance with the exemplary embodiments, the disclosed computer programs can be executed in many exemplary ways, such as an application that is resident in the memory of a device or as a hosted application that is being executed on a server and communicating with the device application or browser via a number of standard protocols, such as TCP / IP, HTTP, XML, SOAP, REST, JSON and other sufficient protocols. The disclosed computer programs can be written in exemplary programming languages that execute from memory on the device or from a hosted server, such as BASIC, COBOL, C, C++, Java, Pascal, or scripting languages such as JavaScript, Python, Ruby, PHP, Perl, or other suitable programming languages.

[0113] As used herein, the terms “computing device” and “entities” should be broadly construed and should be understood to be interchangeable. They may include any type of computing device, for example, a server, a desktop computer, a laptop computer, a smart phone, a cell phone, a pager, a personal digital assistant (PDA, e.g., with GPRS NIC), a mobile computer with a smartphone client, or the like.

[0114] As used herein, a user interface is generally a system by which users interact with a computing device. A user interface can include an input for allowing users to manipulate a computing device, and can include an output for allowing the system to present information and / or data, indicate the effects of the user’s manipulation, etc. An example of a user interface on a computing device (e.g., a mobile device) includes a graphical user interface (GUI) that allows users to interact with programs in more ways than typing. A GUI typically can offer display objects, and visual indicators, as opposed to text-based interfaces, typed command labels or text navigation to represent information and actions available to a user. For example, an interface can be a display window or display object, which is selectable by a user of a mobile device for interaction. A user interface can include an input for allowing users to manipulate a computing device, and can include an output for allowing the computing device to present information and / or data, indicate the effects of the user’s manipulation, etc. An example of a user interface on a computing device includes a graphical user interface (GUI) that allows users to interact with programs or applications in more ways than typing. A GUI typically can offer display objects, and visual indicators, as opposed to text-based interfaces, typed command labels or text navigation to represent information and actions available to a user. For example, a user interface can be a display window or display object, which is selectable by a user of a computing device for interaction. The display object can be displayed on a display screen of a computing device and can be selected by and interacted with by a user using the user interface. In an example, the display of the computing device can be a touch screen, which can display the display icon. The user can depress the area of the display screen where the display icon is displayed for selecting the display icon. In another example, the user can use any other suitable user interface of a computing device, such as a keypad, to select the display icon or display object. For example, the user can use a mouse, a track pad, a track ball, or arrow keys for moving a cursor to highlight and select the display object.

[0115] The display object can be displayed on a display screen of a mobile device and can be selected by and interacted with by a user using the interface. In an example, the display of the mobile device can be a touch screen, which can display the display icon. The user can depress the area of the display screen at which the display icon is displayed for selecting the display icon. Inanother example, the user can use any other suitable interface of a mobile device, such as a keypad, to select the display icon or display object.

[0116] As referred to herein, a computer network may be any group of computing systems, devices, or equipment that are linked together. Examples include, but are not limited to, local area networks (LANs) and wide area networks (WANs). A network may be categorized based on its design model, topology, or architecture. In an example, a network may be characterized as having a hierarchical internetworking model, which divides the network into three layers: access layer, distribution layer, and core layer. The access layer focuses on connecting client nodes, such as workstations to the network. The distribution layer manages routing, filtering, and quality-of-server (QoS) policies. The core layer can provide high-speed, highly-redundant forwarding services to move packets between distribution layer devices in different regions of the network. The core layer typically includes multiple routers and switches.

[0117] The present subject matter may be a system, a method, and / or a computer program product. The computer program product may include a computer readable storage medium (or media) having computer readable program instructions thereon for causing a processor to carry out aspects of the present subject matter.

[0118] The computer readable storage medium can be a tangible device that can retain and store instructions for use by an instruction execution device. The computer readable storage medium may be, for example, but is not limited to, an electronic storage device, a magnetic storage device, an optical storage device, an electromagnetic storage device, a semiconductor storage device, or any suitable combination of the foregoing. A non-exhaustive list of more specific examples of the computer readable storage medium includes the following: a portable computer diskette, a hard disk, a RAM, a ROM, an erasable programmable read-only memory (EPROM or Flash memory), a static random access memory (SRAM), a portable compact disc read-only memory (CD-ROM), a digital versatile disk (DVD), a memory stick, a floppy disk, a mechanically encoded device such as punch-cards or raised structures in a groove having instructions recorded thereon, and any suitable combination of the foregoing. A computer readable storage medium, as used herein, is not to be construed as being transitory signals per se, such as radio waves or other freely propagating electromagnetic waves, electromagnetic waves propagating through a waveguide or other transmission media (e.g., light pulses passing through a fiber-optic cable), or electrical signals transmitted through a wire.

[0119] Computer readable program instructions described herein can be downloaded to respective computing / processing devices from a computer readable storage medium or to an external computer or external storage device via a network, for example, the Internet, a local areanetwork, a wide area network and / or a wireless network, or Near Field Communication. The network may comprise copper transmission cables, optical transmission fibers, wireless transmission, routers, firewalls, switches, gateway computers and / or edge servers. A network adapter card or network interface in each computing / processing device receives computer readable program instructions from the network and forwards the computer readable program instructions for storage in a computer readable storage medium within the respective computing / processing device.

[0120] Computer readable program instructions for carrying out operations of the present subject matter may be assembler instructions, instruction-set-architecture (ISA) instructions, machine instructions, machine dependent instructions, microcode, firmware instructions, statesetting data, or either source code or object code written in any combination of one or more programming languages, including an object oriented programming language such as Java, Smalltalk, C++, Javascript or the like, and conventional procedural programming languages, such as the "C" programming language or similar programming languages. The computer readable program instructions may execute entirely on the user's computer, partly on the user's computer, as a stand-alone software package, partly on the user's computer and partly on a remote computer, or entirely on the remote computer or server. In the latter scenario, the remote computer may be connected to the user's computer through any type of network, including a local area network (LAN) or a wide area network (WAN), or the connection may be made to an external computer (for example, through the internet using an Internet Service Provider). In some embodiments, electronic circuitry including, for example, programmable logic circuitry, field-programmable gate arrays (FPGA), or programmable logic arrays (PLA) may execute the computer readable program instructions by utilizing state information of the computer readable program instructions to personalize the electronic circuitry, in order to perform aspects of the present subject matter.

[0121] Aspects of the present subject matter may be described herein with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the subject matter. It will be understood that each block of the flowchart illustrations and / or block diagrams, and combinations of blocks in a flowchart illustrations and / or block diagrams, can be implemented by computer readable program instructions.

[0122] These computer readable program instructions may be provided to a processor of a computer, special purpose computer, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create means for implementing the functions / actsspecified in the flowchart and / or block diagram block or blocks. These computer readable program instructions may also be stored in a computer readable storage medium that can direct a computer, a programmable data processing apparatus, and / or other devices to function in a particular manner, such that the computer readable storage medium having instructions stored therein comprises an article of manufacture including instructions which implement aspects of the function / act specified in a flowchart and / or block diagram block or blocks.

[0123] The computer readable program instructions may also be loaded onto a computer, other programmable data processing apparatus, or other device to cause a series of operational steps to be performed on the computer, other programmable apparatus, or other device to produce a computer implemented process, such that the instructions which execute on the computer, other programmable apparatus, or other device implement the functions / acts specified in the flowchart and / or block diagram block or blocks.

[0124] The description may illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various embodiments of the present subject matter. In this regard, each block in the flowchart or block diagrams may represent a module, segment, or portion of instructions, which comprises one or more executable instructions for implementing the specified logical function(s). In some alternative implementations, the functions noted in the block may occur out of the order noted herein. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality involved. It will also be noted that each block of the block diagrams and / or flowchart illustration, and combinations of blocks in the block diagrams and / or flowchart illustration, can be implemented by special purpose hardware-based systems that perform the specified functions or acts or carry out combinations of special purpose hardware and computer instructions.

[0125] Samples

[0126] The sample may be from a subject suspected of having an insect bite. The biological sample may also be from a subject with an ambiguous diagnosis in order to clarify the diagnosis.

[0127] The sample may also be obtained for the purpose of prognosis, i.e., determining the course of the disease and selecting primary treatment options. The sample may also be evaluated to select or monitor therapy, selecting likely responders in advance from non-responders or monitoring response in the course of therapy. In addition, the sample may be evaluated as part of post-treatment ongoing surveillance of patients.

[0128] Samples may be obtained using any of a number of methods in the art. Examples of biological samples comprising potential cancer cells include those obtained from excised skinbiopsies, such as punch biopsies, shave biopsies, core needle biopsies, fine needle aspirates (FNA), or surgical excisions; or biopsy from non- cutaneous tissues such as lymph node tissue, mucosa, other embodiments. Representative biopsy techniques include, but are not limited to, excisional biopsy, incisional biopsy, pinch biopsy, forceps biopsy, needle biopsy, or surgical biopsy.

[0129] A sample may also be a sample from a mucosal surface, blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, white blood cells, circulating tumor cells isolated from blood, free DNA isolated from blood, and the like), sputum, saliva, lymph and tongue tissue, cultured cells, e.g., primary cultures, explants, and transformed cells, stool, urine, etc. The sample may be a sample from a whole intact tick, a portion of a tick, a pooled collection of ticks. A sample may be from a eukaryotic organism, e.g., an aves (a bird), e.g., a chicken, a duck, a goose, a ground nesting bird, or a turkey; e.g., a felid, e.g., a bobcat, a cougar, a domestic cat, a lynx, or a mountain lion; e.g., a mammal, e.g., a bear, a camel, a cow, a deer, a dog, a fox, a goat, a horse, a human, a pig, a possum, a rabbit, a raccoon, a sheep, a shrew, or a wild boar; e.g., a primate e.g., human or monkey; a rodent, e.g., a chipmunk, a guinea pig, a mouse, a North American deer mouse, a rat, a squirrel, a vole, or a white-footed mouse. It may be from a domestic animal, a farm animal, or a wild animal. It may be a sewage sample, a soil sample, or a water sample.

[0130] A sample may be treated with a fixative such as formaldehyde and embedded in paraffin (FFPE) and sectioned for use in the methods of the invention. Alternatively, fresh or frozen samples may be used. The sample may be fixed, e.g., in alcoholic solutions such as 100% ethanol or 3:1 methanol: acetic acid. Typically, biological samples, once obtained, are processed prior to nucleic acid analysis using standard methods known in the art. In some embodiments, such processing includes protease treatment and additional fixation in an aldehyde solution such as formaldehyde. In other embodiments, the DNA and / or RNA may be extracted using a commercially available kit such as kits from Qiagen (Germantown, MD), e.g. QIAamp DNA Micro Kit or QIAsymphony SP; Promega (Madison, WI) e.g. Wizard® Genomic DNA Purification Kit; or Zymo Research (Irvine, CA) (see the Example section).

[0131] Polynucleotide Sequence Amplification and Determination

[0132] In many instances, it is desirable to amplify a nucleic acid sequence using any of several nucleic acid amplification procedures which are well known in the art. Specifically, nucleic acid amplification is the chemical or enzymatic synthesis of nucleic acid copies which contain a sequence that is complementary to a nucleic acid sequence being amplified (template). Preferably, the samples are amplified by PCR. The PCR process is well known in the art and isthus not described in detail herein. For a review of PCR methods and protocols, see, e.g., Innis et al., eds., PCR Protocols, A Guide to Methods and Application, Academic Press, Inc., San Diego, Calif. 1990; U. S. Pat. No. 4,683,202 (Mullis); which are incorporated herein by reference in their entirety. PCR reagents and protocols are also available from commercial vendors, such as Roche Molecular Systems. PCR may be carried out as an automated process with a thermostable enzyme. In this process, the temperature of the reaction mixture is cycled through a denaturing region, a primer annealing region, and an extension reaction region automatically. Machines specifically adapted for this purpose are commercially available.

[0133] Alternatively, other amplification techniques include Ligase Chain Reaction (LCR), Polymerase Ligase Chain Reaction, Gap-LCR, Repair Chain Reaction, 3 SR, nucleic acid sequence-based amplification (NASBA), Strand Displacement Amplification (SDA), Transcription Mediated Amplification (TMA), and QP-amplification. The methods and kits of the invention may use any nucleic acid amplification or detection methods known to one skilled in the art, such as those described in U. S. Pat. Nos. 5,525,462 (Takarada et al.) 6,114,117 (Hepp et al.) 6,127,120 (Graham et al.); 6,344,317 (Urnovitz); 6,448,001 (Oku); 6,528,632 (Catanzariti et al.); and PCT Pub. No. WO 2005 / 111209 (Nakajima et al.y all of which are incorporated herein by reference in their entirety.

[0134] In some embodiments, the nucleic acids may be amplified by PCR amplification using methodologies known to one skilled in the art. One skilled in the art will recognize, however, that amplification can be accomplished by other known methods, such as LCR, QP-replicase amplification, rolling circle amplification, transcription amplification, self-sustained sequence replication, NASBA, each of which provides sufficient amplification. Branched-DNA technology may also be used to qualitatively demonstrate the presence of a sequence of the technology which may quantitatively determine the amount of a particular genomic sequence in a sample. Nolte reviews branched-DNA signal amplification for direct quantitation of nucleic acid sequences in clinical samples (Nolte, 1998, Adv. Clin. Chem. 33:201-235).

[0135] High Throughput and Single Molecule Sequencing Technology

[0136] Suitable next generation sequencing technologies are widely available. Examples include the 454 Life Sciences platform (Roche, Branford, CT) (Margulies etal. 2005 Nature, 437, 376-380); Illumina’s Genome Analyzer, Illumina’s MiSeq System, Illumina’s NextSeq System, Illumina’s MiniSeq System (Illumina, San Diego, CA; Bibkova et al., 2006, Genome Res. 16, 383-393; U. S. Pat. Nos. 6,306,597 (Macevicz), 7,232,656 (Balasubramanian et al.), 7,598,035 (Macevicz)); or DNA Sequencing by Ligation, SOLiD System (Applied Biosystems / Life Technologies; U. S. Pat. Nos. 6,797,470, 7,083,917, 7,166,434, 7,320,865, 7,332,285, 7,364,858,and 7,429,453 (Barany etal.y, or the Helicos True Single Molecule DNA sequencing technology (Harris et al., 2008 Science, 320, 106-109; U. S. Pat. Nos. 7,037,687 (Williams et al.), 7,169,560 (Lapidus et al.), 7,645,596 (Williams et al.), 7,769,400 (Harris)), the single molecule, real-time (SMRT™) technology of Pacific Biosciences, and sequencing (Soni and Meller, 2007, Clin. Chem. 53, 1996-2001) which are incorporated herein by reference in their entirety. These systems allow the sequencing of many nucleic acid molecules isolated from a specimen at high orders of multiplexing in a parallel fashion (Dear, 2003, Brief Funct. Genomic Proteomic, 1(4), 397-416 and McCaughan and Dear, 2010, J. Pathol., 220, 297-306). Each of these platforms allow sequencing of clonally expanded or non-amplified single molecules of nucleic acid fragments. Certain platforms involve, for example, (i) sequencing by ligation of dye-modified probes (including cyclic ligation and cleavage), (ii) pyrosequencing, (iii) targeted next-generation sequencing from bisulfite treated DNA, and (iv) single-molecule sequencing.

[0137] Pyrosequencing is a nucleic acid sequencing method based on sequencing by synthesis, which relies on detection of a pyrophosphate released on nucleotide incorporation. Sequencing by synthesis involves synthesizing, one nucleotide at a time, a DNA strand complimentary to the strand whose sequence is being sought. Study nucleic acids may be immobilized to a solid support, hybridized with a sequencing primer, incubated with DNA polymerase, ATP sulfurylase, luciferase, apyrase, adenosine 5' phosphsulfate and luciferin. Nucleotide solutions are sequentially added and removed. Correct incorporation of a nucleotide releases a pyrophosphate, which interacts with ATP sulfurylase and produces ATP in the presence of adenosine 5' phosphosulfate, fueling the luciferin reaction, which produces a chemiluminescent signal allowing sequence determination. Machines for pyrosequencing are available from Qiagen, Inc. (Valencia, CA). An example of a system that can be used by a person of ordinary skill based on pyrosequencing may involve the following steps: ligating an adaptor nucleic acid to a study nucleic acid and hybridizing the study nucleic acid to a bead; amplifying a nucleotide sequence in the study nucleic acid in an emulsion; sorting beads using a picoliter multiwell solid support; and sequencing amplified nucleotide sequences by pyrosequencing methodology (e.g., Nakano etal., 2003, J. Biotech. 102, 117-124). Such a system can be used to exponentially amplify amplification products generated by a process described herein, e.g., by ligating a heterologous nucleic acid to the first amplification product generated by a process described herein.

[0138] Another next-generation sequencing (NGS) is a nucleic acid sequencing method based on sequencing by synthesis, where fluorescently labeled deoxyribonucleotide triphosphates (dNTPs) catalyzed by DNA polymerase are incorporated into a DNA temple through cycles of DNA synthesis and nucleotides are identified by fluorophore excitation at each incorporation step.NGS allows this process to take place in a multiplex reaction across millions of DNA fragments in parallel. Generally, sequencing by synthesis involves synthesizing, one nucleotide at a time, a DNA strand complimentary to the strand whose sequence is being sought. Study nucleic acids may be immobilized to a solid support, hybridized with a sequencing primer, and incubated with DNA polymerase in the presence of fluorescently labeled dNTPS. After each cycle, the image is scanned and the emission wavelength and intensity are recorded and used to identify the base incorporated. This process is repeated multiple times to create a specific read length of bases.

[0139] Certain single-molecule sequencing embodiments are based on the principal of sequencing by synthesis and utilize single-pair Fluorescence Resonance Energy Transfer (single pair FRET) as a mechanism by which photons are emitted as a result of successful nucleotide incorporation. The emitted photons often are detected using intensified or high sensitivity cooled charge-couple-devices in conjunction with total internal reflection microscopy (TIRM). Photons are only emitted when the introduced reaction solution contains the correct nucleotide for incorporation into the growing nucleic acid chain that is synthesized as a result of the sequencing process. In FRET based single-molecule sequencing or detection, energy is transferred between two fluorescent dyes, sometimes polymethine cyanine dyes Cy3 and Cy5, through long-range dipole interactions. The donor is excited at its specific excitation wavelength and the excited state energy is transferred, non-radiatively to the acceptor dye, which in turn becomes excited. The acceptor dye eventually returns to the ground state by radiative emission of a photon. The two dyes used in the energy transfer process represent the "single pair", in single pair FRET. Cy3 often is used as the donor fluorophore and often is incorporated as the first labeled nucleotide. Cy5 often is used as the acceptor fluorophore and is used as the nucleotide label for successive nucleotide additions after incorporation of a first Cy3 labeled nucleotide. The fluorophores generally are within 10 nanometers of each other for energy transfer to occur successfully.

[0140] An example of a system that can be used based on single-molecule sequencing generally involves hybridizing a primer to a study nucleic acid to generate a complex; associating the complex with a solid phase; iteratively extending the primer by a nucleotide tagged with a fluorescent molecule; and capturing an image of fluorescence resonance energy transfer signals after each iteration (e.g., Braslavsky et al., PNAS 100(7): 3960-3964 (2003); U. S. Pat. No.7,297,518 (Quake et al.) which are incorporated herein by reference in their entirety). Such a system can be used to directly sequence amplification products generated by processes described herein. In some embodiments, the released linear amplification product can be hybridized to a primer that contains sequences complementary to immobilized capture sequences present on a solid support, a bead or glass slide for example. Hybridization of the primer-released linearamplification product complexes with the immobilized capture sequences, immobilizes released linear amplification products to solid supports for single pair FRET based sequencing by synthesis. The primer often is fluorescent, so that an initial reference image of the surface of the slide with immobilized nucleic acids can be generated. The initial reference image is useful for determining locations at which true nucleotide incorporation is occurring. Fluorescence signals detected in array locations not initially identified in the "primer only" reference image are discarded as non-specific fluorescence. Following immobilization of the primer-released linear amplification product complexes, the bound nucleic acids often are sequenced in parallel by the iterative steps of, a) polymerase extension in the presence of one fluorescently labeled nucleotide, b) detection of fluorescence using appropriate microscopy, TIRM for example, c) removal of fluorescent nucleotide, and d) return to step a with a different fluorescently labeled nucleotide.

[0141] The technology described herein may be practiced with digital PCR. Digital PCR was developed by Kalinina and colleagues (Kalinina et al., 1997, Nucleic Acids Res. 25; 1999-2004) and further developed by Vogelstein and Kinzler (1999, Proc. Natl. Acad. Sci. U. S. A. 96; 9236-9241). The application of digital PCR is described by Cantor et al. (PCT Pub. Nos. WO 2005 / 023091A2 (Cantor et al.),' WO 2007 / 092473 A2, (Quake et al which are hereby incorporated by reference in their entirety. Digital PCR takes advantage of nucleic acid (DNA, cDNA or RNA) amplification on a single molecule level, and offers a highly sensitive method for quantifying low copy number nucleic acids. Fluidigm® Corporation offers systems for the digital analysis of nucleic acids.

[0142] In some embodiments, nucleotide sequencing may be by solid phase single nucleotide sequencing methods and processes. Solid phase single nucleotide sequencing methods involve contacting sample nucleic acid and solid support under conditions in which a single molecule of sample nucleic acid hybridizes to a single molecule of a solid support. Such conditions can include providing the solid support molecules and a single molecule of sample nucleic acid in a "microreactor." Such conditions also can include providing a mixture in which the sample nucleic acid molecule can hybridize to solid phase nucleic acid on the solid support. Single nucleotide sequencing methods useful in the embodiments described herein are described in PCT Pub. No. WO 2009 / 091934 (Cantor).

[0143] In certain embodiments, nanopore sequencing detection methods include (a) contacting a nucleic acid for sequencing ("base nucleic acid," e.g., linked probe molecule) with sequence-specific detectors, under conditions in which the detectors specifically hybridize to substantially complementary subsequences of the base nucleic acid; (b) detecting signals from the detectors and (c) determining the sequence of the base nucleic acid according to the signalsdetected. In certain embodiments, the detectors hybridized to the base nucleic acid are disassociated from the base nucleic acid (e.g., sequentially dissociated) when the detectors interfere with a nanopore structure as the base nucleic acid passes through a pore, and the detectors disassociated from the base sequence are detected.

[0144] A detector also may include one or more regions of nucleotides that do not hybridize to the base nucleic acid. In some embodiments, a detector is a molecular beacon. A detector often comprises one or more detectable labels independently selected from those described herein. Each detectable label can be detected by any convenient detection process capable of detecting a signal generated by each label (e.g., magnetic, electric, chemical, optical and the like). For example, a CD camera can be used to detect signals from one or more distinguishable quantum dots linked to a detector.

[0145] The invention encompasses methods known in the art for enhancing the sensitivity of the detectable signal in such assays, including, but not limited to, the use of cyclic probe technology (Bakkaoui et al., 1996, BioTechniques 20: 240-8, which is incorporated herein by reference in its entirety); and the use of branched probes (Urdea etal., 1993, Clin. Chem. 39, 725-6; which is incorporated herein by reference in its entirety). The hybridization complexes are detected according to well-known techniques in the art.

[0146] Reverse transcribed or amplified nucleic acids may be modified nucleic acids. Modified nucleic acids can include nucleotide analogs, and in certain embodiments include a detectable label and / or a capture agent. Examples of detectable labels include, without limitation, fluorophores, radioisotopes, colorimetric agents, light emitting agents, chemiluminescent agents, light scattering agents, enzymes and the like. Examples of capture agents include, without limitation, an agent from a binding pair selected from antibody / antigen, antibody / antibody, antib ody / antibody fragment, antibody / antibody receptor, antibody / protein A or protein G, hapten / anti -hapten, biotin / avidin, biotin / streptavidin, folic acid / folate binding protein, vitamin B12 / intrinsic factor, chemical reactive group / complementary chemical reactive group (e.g., sulfhydryl / maleimide, sulfhydryl / haloacetyl derivative, amine / isotriocyanate, amine / succinimidyl ester, and amine / sulfonyl halides) pairs, and the like. Modified nucleic acids having a capture agent can be immobilized to a solid support in certain embodiments.

[0147] Next generation sequencing techniques may be applied to measure expression levels or count numbers of transcripts using RNA-seq or whole transcriptome shotgun sequencing. See, e.g., Mortazavi et al. 2008 Nat Meth 5(7) 621-627 or Wang et al. 2009 Nat Rev Genet 10(1) 57-63. Nucleic acids in the invention may be counted using methods known in the art. In one embodiment, NanoString’s nCounter® system may be used (Seattle, WA). Geiss et al. 2008 NatBiotech 26(3) 317-325; U. S. Pat. No. 7,473,767 (Dimitrov). In addition, NanoString’s Digital Spatial Profiling (DSP) platform may be used for nucleic acid or protein detection. Blank et al., 2018 Nature Medicine 24 1655-1661; Amaria et al., 2018 Nature Medicine 24 1649-1654. Alternatively, Fluidigm’s Dynamic Array system may be used (South San Francisco, CA). Byrne et al. 2009 PLoS ONE 4 e7118; Helzer et al. 2009 Can Res 69 7860-7866. For reviews, see also Zhao et al. 2011 Sci China Chem 54(8) 1185-1201 and Ozsolak and Milos 2011 Nat Rev Genet 12 87-98.

[0148] Compositions and Kits

[0149] The invention provides compositions and kits identifying the pathogens, the ticks, and / or the blood meal hosts described herein using reagents specific for the nucleic acids specific for the polynucleotides. Kits for carrying out the diagnostic assays of the invention typically include, in suitable container means, (i) a forward and reverse primer for the marker polynucleotides of the invention; and (ii) an extension primer that comprises nucleic acid sequence that specifically binds to the marker polynucleotides of the invention. The kits may include several polynucleotide sequences encoding biomarkers disclosed herein, e.g., specific nucleic acids. In one embodiment the nucleic acids in the kit are the forward and reverse PCR primers and the extension primers for the biomarkers disclosed herein. The container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe and / or other container into which a first nucleic acid specific for one of the polynucleotides of the present invention may be placed and / or suitably aliquoted. Where a second and / or third and / or additional component is provided, the kit will also generally contain a second, third and / or another additional container into which this component may be placed. Alternatively, a container may contain a mixture of more than one nucleic acid reagent, each reagent specifically binding a different marker in accordance with the present invention. The kits of the present invention will also typically include means for containing nucleic acid probes in close confinement for commercial sale. Such containers may include injection and / or blow-molded plastic containers into which the desired vials are retained.

[0150] The kits may further comprise positive and negative controls, as well as instructions for the use of kit components contained therein, in accordance with the methods of the present invention. In some embodiments, the kits may include DNA / RNA extraction reagents, see e.g.,

[0100] ,

[0151] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Preferred methods, devices, and materials are described, although any methods andmaterials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure. All references cited herein are incorporated by reference in their entirety.

[0152] The following Examples further illustrate the disclosure and are not intended to limit the scope. In particular, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.6. EXAMPLES

[0153] GENERAL ASSAY METHOD

[0154] Assay Design

[0155] The procedures of PCT publication WO2016 / 172579 published 27 Oct. 2016 and W02017 / 049180 published 23 March 2017 (Agena Biosciences, Inc., San Diego, CA)Each assay consists of three primers, two PCR primers and one single base extension primer. All amplicons are under 150bp in length to ensure amplification success and mass tags are added to the 5' end of the primer to move unincorporated PCR primers out of the analytical mass window. Extension probe design also has few requirements. First, the mass of the extended products must be sufficiently spaced by mass to ensure no conflicts between assays. Second, multiplexing is based on mutant allele (low-abundance variant) hence in one reaction only minor allele variants (low-abundance variants) with an identical nucleotide incorporated during the single base extension are multiplexed together. No concern is taken to the major allele (high-abundance variant) as long as it is different from the minor allele. The extension product for the major allele (high-abundance variant) provides a control for distinguishing a wild type result from a failed assay. Optional control assays, for example, may include capture controls to verify that the bead capture, cleaning, and elution steps were successful.

[0156] PCR amplification

[0157] In a typical experiment PCR is carried out in a total volume of 20pL with lOpL of DNA template supplemented with lOpL of a master mix consisting of lx PCR Buffer supplemented with 1 mM MgCh, 125pM dNTPs, 0.125U Uracil-DNA glycosylase, 4U Taq polymerase, and lOOnM of each PCR primer. Reactions are initially incubated at 30°C for 10 minutes followed by 94°C for 2 minutes. 45 cycles of PCR were performed at 94 °C for 30 seconds, 56°C for 30 seconds, and 72°C for 1 minute. The PCR is completed with a final incubation of 5 minutes at 72°C. 5pL of amplified products are conditioned with the addition of2pL of 0.5U shrimp alkaline phosphatase (SAP) in 0.24x SAP buffer in a total volume of 7pL for 40 minutes at 37°C followed by SAP enzyme denaturation for 10 minutes at 85°C.

[0158] Single base extension

[0159] Single base extension is performed by adding 2pL of a mastermix consisting of 0.2x extension buffer, 5.56pM of the minor allele variant nucleotide and different concentrations of the major allele variant nucleotides ranging from 0.03-1.25 pM, extension primers at various concentrations, and 0.14U iPLEX® Pro enzyme. Single base extension reactions are performed in a total volume of 9pL. Reaction parameters include an initial incubation at 94°C for 30 seconds followed by 40 cycles at 94°C for 5 seconds with five nested cycles of 52°C for 5 seconds then 80°C for 5 seconds. The single base extension is completed with an incubation at 72°C for 3 minutes.

[0160] Capture and data acquisition

[0161] 1. Desalting: Add HPLC-grade water to each well of the reaction plate using a 12-channel multipipettor. iPLEX panels typically require 16 pL per well for 384-well plates, and 30 pL or more for 96-well plates. 2. Seal the plate and centrifuge the plate at 3200 x g for 1 minute.3. Process the plate on the MassARRAY System with Chip Prep Module (CPM) preloaded with resin beads or MassARRAY Nanodispenser RS 1000 preloaded with resin beads and MassARRAY Analyzer.

[0162] In CPM or RS 1000 instrument, the supernatant is dispensed onto a Spectrochip® 11 solid support / chip. Data is acquired via MALDI-TOF mass spectrometry using the MassARRAY® 4 instrument.

[0163] SPECIFIC PROTOCOLS FOR DNA OR RNA EXTRACTION FROM SAMPLES

[0164] DNA or RNA extraction from arthropods, arthropod-bite wounds, synovial fluid, or whole blood.

[0165] This is the specific protocol used to extract DNA from a single arthropod or pooled sample of arthropods. Key reagents and instruments:ZymoBIOMICS (Zymo Research, Irvine, CA) 96 Magbead DNA KIT( No Lysis matrix) ZR BashingBead (Zymo Research) lysis tubes (0.1 & 2.0 mm)BERTIN Precellys Evolution Touch Homogenizer or Bead (Bertin Technologies, Montigny-le-Bretonneux, FRANCE)

[0166] Total DNA / RNA extractionZymoBIOMICS MagBead DNA / RNA kit or other total nucleic acid extraction kit.ZymoBIOMICS BashingBeads lysis rack (0.1 & 0.5mm)Add 750 pl DNA / RNA Shield™ to a sample (see table below) and mix. If a sample is already collected in DNA / RNA Shield™, proceed to step 2.Sample Type: arthropod (e.g. ticks, mosquitoes), whole blood, tissue, swabs, other body fluidsFor complete microbial lysis, perform mechanical homogenization via bead beating (recommended: ZR BashingBead™ Lysis Tubes (0.1 & 0.5 mm, S6012-50) or ZymoBIOMICS BashingBead™ Lysis Rack (0.1 & 0.5 mm, S6002-96-7).Transfer entire liquid sample into a lysis tube / rack and secure in a high-speed bead beater ( Bertin Precellys, etc.). Process 1 at maximum speed for > 5 minutes.Centrifuge and transfer up to 200 pl of the supematant2 into a nuclease-free tube (not provided).5. Add 10 pl Proteinase K for every 200 pl sample. Mix and incubate at room temperature (20-30°C) for 30 minutes.Add 200 pl (1 volume) DNA / RNA Lysis Buffer to 200 pl sample and mix welll.Add 400 pl ethanol (95-100%) to the sample and mix welll.Add 30 pl ZymoBIOMICS™ MagBinding Beads and mix welll for 20 minutes. Important: ZymoBIOMICS™ MagBinding Beads settle quickly, ensure that beads are kept in suspension while dispensing.Transfer the plate / tube to the magnetic stand2 until beads have pelleted, then aspirate3 and discard the cleared supernatant.Add 500 pl MagBead DNA / RNA Wash 1 and mix welll. Pellet the beads and discard the supernatant.Add 500 pl MagBead DNA / RNA Wash 2 and mix welll. Pellet the beads and discard the supernatant.Add 500 pl ethanol (95-100%) and mix welll. Pellet the beads2,3 and discard the supernatantRepeat step 7.Dry the beads for 10 minutes or until dryTo elute DNA / RNA from the beads, add 50 pl ZymoBIOMICS™ DNase / RNase-Free Water and mix welll for 5 minutes.Transfer the plate / tube to the magnetic stand until beads have pelleted, then aspirate3 and dispense the eluted DNA / RNA to a new plate / tube.The eluted DNA / RNA can be used immediately or stored frozen.

[0167] Arthropod DNA extraction

[0168] Arthropod lysis and DNA extraction:o Add up to 10 ticks / mosquitoes in a ZR BashingBead™ Lysis Tube (2.0 mm) (Cat. No.S6003-50) with 750 pl of ZymoBIOMICS™ Lysis Solution (Zymo Research).o Secure in a Precellys Evolution Homogenizer fitted with a 2 ml tube holder assembly and process at 9000rpm with three cycles of 1 minute beating, and 2 minutes resting.o At high-speed 12,000 rpm spin down beads and residues for 10 minutes, then transfer supernatant to a well on a 96-deep well plate.o Run DNA purification protocol “D4302_ZymoBIOMICS DNA KingFisher Flex vl” (Appendix 1) on KingFisher™ Flex instrument (Thermo Fisher Scientific, Waltham, MA) o Store extracted DNA in -20° C freezer.o Run MassARRAY (Agena Research) protocol with four reactions as PHI, PH2, PH3 and PH4.

[0169] DNA Extraction from swabs collected from an arthropod-bite wound

[0170] This is the protocol used for swabs collected from bite wounds without the arthropod.o Break buccal swab into 200ul of PBS solution in a ZR BashingBead™ Lysis Tube with 0.1mm and 0.5mm beads and tighten the cap.o Add 750 pl of ZymoBIOMICS™ Lysis Solution, then assemble the tube onto the Beadbeater tube tray, and run bead beating protocol.o Secure in a Precellys Evolution Homogenizer fitted with a 2 ml tube holder assembly and process at 9000rpm with three cycles of 1 minute beating, and 2 minutes resting.o At high-speed 12,000 rpm spin down beads and residues for 10 minutes, then transfer supernatant to a well on a 96-deep well plate.o Run DNA purification protocol “D4302_ZymoBIOMICS DNA King Fisher Flex vl” (Appendix 1) on King Fisher Flex instrument.o Store extracted DNA in -20° C freezer.o Run MassARRAY protocol with four reactions PHI, PH2, PH3 and PH4.

[0171] Extract pathogen and / or arthropod DNA from whole blood or synovial fluids

[0172] This is the protocol used to extract pathogen and / or arthropod DNA from whole blood or synovial fluids. Key reagents and instruments:o HostZERO Microbial DNA kit (Zymo Research)o RBC Lysis buffer #R1022-2- 100 (Zymo Research)o ZR BashingBead Lysis Tube (0.1 & 0.5 mm)

[0173] Host DNA depletion with the whole blood sampleo In a 15 ml conical tube, add 9 ml RBC Lysis Buffer into 3 ml of blood.o Mix by inverting and incubate for 5min at room temperature.o Centrifuge at 2,000xg for 10 minutes to pellet cells, if available, centrifugation at 8,000- 10,000xg for 10 minutes is preferred.o Carefully remove and discard the supernatant without disturbing the pellet.o Resuspend the pellet in 200 ul of PBS and transfer to a clean microcentrifuge tube. o Add 1ml host DNA Depletion Solution (Zymo Research) to 200ul of PBS suspension. o Rotate sample for 15 min using end-over-end rotation at room temperature (20-30° C) o Centrifuge the tube at 10,000xg for 5 minutes.o Carefully remove and discard the supernatant without disturbing the pellet.o Add 100 ul of Microbial Selection Buffer (Zymo Research) to the tube and resuspend the pellet.o Add 1 ul of Microbial Selection Enzyme (Zymo Research) to the suspension, vortex briefly to mix.o Incubate the tube at 37° C for 30 minutes.o Add 20ul of Proteinase K to the sample and vortex for at least 10 seconds, incubate at 55° C for 10 minutes.o Add 100 ul of DNA / RNA Shield (2X Concentrate) (Zymo Research) to the sample and vortex for 10 second. Incubate at room temperature for 5 minutes.o Proceed to Microbial DNA isolation or store samples at -20° C.

[0174] Microbial DNA isolation

[0175] This is the procedure for microbial DNA isolation.o Add all suspended samples to a ZR BashingBead Lysis Tube (0.1& 0.5mm). add 750 ul of ZymoBIOMICS Lysis Solution to the tube and cap tightly.o Secure in a Bertin Evolution Homogenizer tube holder and run beating program (3 cycle of 1 min 9000rpm and 2 min rest).o Centrifuge the ZR BashingBead Lysis Tube (0.1 &0.5mm) in a microcentrifuge at >10,000xg for 1 minute.o Transfer 400-600 ul of supernatant to a collection Tube, add 1200 ul of ZymoBIOMICS DNA Binding Buffer (Zymo Research) to the supernatant and mix well.o Transfer the supernatant to Zymo-Spin IC-Z Column (Zymo Research) in a collection Tubes and centrifuge. Discard the flow-through.o Add 400 ul of ZymoBIOMICS DNA Wash Buffer 1 (Zymo Research) to the column in a new collection tube and centrifuge. Discard the flow-through.o Add 700 ul of ZymoBIOMICS DNA Wash Buffer 2 (Zymo Research) to the column and centrifuge. Discard the flow-through.o Add 200 ul of ZymoBIOMICS Wash Buffer 2 and centrifuge the column for 1 minute to ensure complete removal of the wash buffer. Then carefully transfer the column into a clean microcentrifuge tube.o Add 20 ul of ZymoBIOMICS DNAase / RNase-free water (Zymo Research) directly to the column matrix and incubate for 5 minutes.o Centrifuge at full speed for 1 minute to elute the DNA.o Store DNA in -20° C freezer.

[0176] PCR AMPLIFICATION AND EXTENSION WITH MASSARRAY ® PROTOCOL

[0177] PCR primer pools are mixed with individuals to a final concentration of 1 µM. Extension primer pools are mixed with individual oligo according to its molecular weight, starting from 4200 Daltons at 5 µM to 9000 Daltons at 15 µM.

[0178] The quantities below for the PCR protocol are meant for 32 samples, 1 µl of DNA sample is added into each well, total reaction volume is 5 µl.

[0179] Shrimp alkaline phosphatase (SAP) treatment after PCR: 2 µl of SAP Master Mix is added into each PCR well.

[0180] After SAP treatment, 2 µl of Extension Master Mix is added into each PCR well.

[0181] The final 9 µl of products are loaded onto a MassARRAY instrument.

[0182] MALDI TOP AND DATA ANALYSIS.

[0183] FIG. 1 shows a summary slide of the pathogens detected, arthropod identified, and blood meal sources from a series of experiments on 116 ticks. FIG. 4-6, FIG. 8-10, and FIG. 13-22 show the results of the pathogen identification analysis. FIG. 2-3, FIG. 7, and FIG. 11-12 show the results for the arthropod identification analysis. FIG. 22-FIG. 30 shows the results for the host (blood meal) identification analysis.

[0184] Specimens:Typical biological samples include synovial fluid, erythema migrans tissue biopsy, cerebrospinal fluid (CSF), whole blood, arthropod (arthropod ID) and host ID.

[0185] ReactionsWell 1: Bacterial / viral + Arthropod ID + Mammal host IDWell 2: virus targets

[0186] Arthropod e.g., Tick Surveillance panel features:Low input: 3 ul of DNA extracted from single or pooled ticksHigh multiplex: 40 reactions in each wellHigh sensitivity: MassARRAY = PCR + MS / MALDI TOFLowest cost per target

[0187] In one set of experiments several hundred samples were tested. Pathogens detected: Anaplasma phagocy tophilum, Babesia microti, Borrelia burgdorferi, Borrelia miyamotoi, Borrelia species, Ehrlichia chaffeensis, Ehrlichia ewingii, Panola Mountain Ehrlichia (PME), Ehrlichia species, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia species, Plasmodium species(sp.), Plasmodium falciparum, Plasmodium knowlesi, Plasmodium ovale, and Plasmodium vivax.

[0188] Arthropod species identified: Amblyomma americanum (Lone star tick), Amblyomma maculatum (Gulf coast tick), Dermacentor variabilis (American dog tick), Ixodes scapularis (deer tick), and Ixodes pacificus (Western black-legged tick)Tick blood meals identified: bird, cat, deer, domestic pig, felids, human, mice, rabbit, shrew, squirrel, vole, and wild boar (feral hogs).

[0189] An example of a tick-borne disease clinical panel

[0190] The panel detected most of CDC-listed tick-borne pathogens with one assay. The panel would assay for human specimens: whole blood, tick-bitten wound, synovial and spinal fluid would be used for patients who experienced: suspected tick-bitten, fever / chills, persistent fever, rash, muscle aches, and joint pains.

[0191] The results are used to diagnose: Lyme disease, babesiosis, ehrlichiosis, malaria (P. falciparum, P. knowlesi, P. ovale, or P. vivax.), rickettsiosis, Rocky Mountain Spotted Fever, anaplasmosis, tick-borne relapsing fever and / or tularemia

[0192] Tick-borne disease clinical panel features:

[0193] The highest sensitivity: cut-edge technology to detect 1-10 bacteria pathogen from 1 mL of human whole blood.

[0194] Quick turn-around-time: a response in less 24 hours after tick bite. Quickest positive detection time upon tick bites, current antibodies-based diagnostic technology needs 7 days incubation time, often delaying patient treatment. The methods disclosed herein analyze 10, 15, 20 or more tick-borne pathogens within one assay. In contrast, many current PCR-based assays only test one pathogen at a time.7. GENERALIZED STATEMENTS OF THE DISCLOSURE

[0195] The following numbered statements provide a general description of the disclosure and are not intended to limit the appended claims.

[0196] Statement 1: A method of identifying (i) an arthropod-borne pathogen, (ii) an arthropod species, and (iii) a blood meal host from a single biological sample, the methodcomprising: (a) amplifying nucleic acids in the biological sample with a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three arthropod species, and (iii) PCR primers specific for at least three blood meal hosts; (b) performing single base extension with a set of extension primers on the amplified products to generate a set of extended products; and (c) analyzing the set of extended products so as to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).

[0197] Statement 2: The method of Statement 1, wherein the arthropod-borne pathogen is a tick-borne pathogen and the arthropod species is a tick.

[0198] Statement 3: The method of Statement 1, wherein the arthropod-borne pathogen is a mosquito-borne pathogen and the arthropod species is a mosquito.

[0199] Statement 4: The method of Statement 1, wherein the arthropod species is a biting midge, a sandfly, or a tsetse fly.

[0200] Statement 5: The method of Statement 1, wherein the arthropod species is a flea, a louse, or a triatomine bug. The arthropod species may be two species e.g., mosquitoes and ticks; ticks and fleas or lice; triatomine bugs and ticks; ticks and flies (biting midges, sandflies, tsetse flies); mosquitoes and flies; mosquitoes and fleas or lice; or three species, ticks, mosquitoes, and fleas / lice; ticks, mosquitoes, and flies.

[0201] Statement 6: The method of any of Statements 1-5, wherein the set of PCR primers comprise (i) PCR primers specific for at least fifteen arthropod-borne pathogens.

[0202] Statement 7: The method of any of Statements 1-5, wherein the set of PCR primers comprise specific for at least twenty arthropod-borne pathogens.

[0203] Statement 8: The method of any of Statements 1-7, wherein the set of PCR primers comprise (ii) PCR primers specific for at least five species of arthropods.

[0204] Statement 9: The method of any of Statements 1-8, wherein the set of PCR primers comprise (iii) PCR primers specific for at least ten blood meal hosts.

[0205] Statement 10: The method of any of Statements 1-9, wherein the arthropod-borne pathogen is a bacterium, a fungus, a protozoan, or a virus.

[0206] Statement 11: The method of Statement 10, wherein the arthropod-borne pathogen is a bacterium and the PCR primers specific for at least ten arthropod-borne pathogens hybridize with a 16S and / or an 18S ribosomal nucleic acid of the bacterium.

[0207] Statement 12: The method of Statement 10, wherein the tick-borne pathogen is a bacterium and the bacterium is an Anaplasma, a Babesia, a Bor re Ha, an Ehrlichia, a Francisella, a Rickettsia, or a Yersinia.

[0208] Statement 13: The method of Statement 13, wherein the Anaplasma, the Babesia, the Bartonella, the Borrelia, the Coxiella, the Ehrlichia, the Francisella, the Rickettsia, or the Yersinia are Anaplasma bovis, Anaplasma caudatum, Anaplasma centrale, Anaplasma marginale, Anaplasma mesaenterum, Anaplasma odocoilei, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma platys, Bartonella bacilliformis, Bartonella elizabethae, Bartonella henselae, Bartonella quintana, Bartonella vinsonii, Borrelia afzelii, Borrelia americana, Borrelia anserina, Borrelia bissettiae, Borrelia burgdorferi, Borrelia californiensis, Borrelia carolinensis, Borrelia coriaceae, Borrelia garinii, Borrelia hermsii, Borrelia kurtenbachii, Borrelia lanei, Borrelia lonestari, Borrelia lusitaniae, Borrelia mayonii, Borrelia miyamotoi, Borrelia parkeri, Borrelia theileri, Borrelia turicatae, Borrelia valaisiana, Coxiella burnetii, Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia mineirensis, Ehrlichia muris, Ehrlichia muris eauclairensis, Ehrlichia muris-like agent, Ehrlichia ruminantium, Ehrlichia sp., Francisella tularensis, Panola Mountain Ehrlichia, Rickettsia africae (aeschlimannii ), Rickettsia akari, Rickettsia amblyommatis, Rickettsia australis, Rickettsia bellii, Rickettsia CA6269, Rickettsia conorii, Rickettsia felis, Rickettsia heilongjiangensis, Rickettsia helvitica, Rickettsia honei, Rickettsia japonica, Rickettsia massiliae, Rickettsia monacensis, Rickettsia montanesis, Rickettsia parkeri, Rickettsia philippi (strain 364D), Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia sibirica, Rickettsia slovaca, Rickettsia typhi, or Yersinia pestis.

[0209] Statement 14: The method of Statement 10, wherein the arthropod-borne pathogen is a protozoan, and the protozoan is a Babesia, a Cytauxzoon, a Leishmania, a Plasmodium, a Theileria, or a Trypanosome.

[0210] Statement 15: The method of Statement 14, wherein the Babesia is Babesia beliceri, Babesia bigemina, Babesia bovis, Babesia caballi, Babesia canis, Babesia catia, Babesia conradae, Babesia crassa, Babesia divergens, Babesia duncani, Babesia felis, Babesia foliatad, Babesia taylori, Babesia gibsoni, Babesia hongkongensis, Babesia jakimovi, Babesia lengau, Babesia major, Babesia microti, Babesia motasi, Babesia occultans, Babesia orientalis, Babesia ovata, Babesia ovis, Babesia perroncitoi, Babesia presentii, Babesia rossi, Babesia trautmanni, Babesia Venator um, or Babesia vogeli.

[0211] Statement 16: The method of Statement 14, wherein the Cytauxzoon is Cytauxzoon banethi, Cytauxzoon europaeus, Cytauxzoon felis. Cytauxzoon manul or Cytauxzoon otrantorum.

[0212] Statement 17: The method of Statement 14, wherein the Leishmania is Leishmania aethiopica, Leishmania amazonensis, Leishmania archibaldi, Leishmania braziliensis, Leishmania colombiensis, Leishmania donovani, Leishmania garnhami, Leishmania guyanensis, Leishmania infantum, Leishmania killicki, Leishmania lainsoni, Leishmania lindenbergi,Leishmania major, Leishmania mexicana, Leishmania naiffi, Leishmania panamensis, Leishmania peruviana, Leishmania pifanoi, Leishmania shawi, Leishmania tropica, or Leishmania venezuelensis.

[0213] Statement 18: The method of Statement 14, wherein the Plasmodium is Plasmodium falciparum, Plasmodium knowlesi, Plasmodium malariae, Plasmodium ovale curtisi, Plasmodium ovale wallikeri, Plasmodium sp., o Plasmodium vivax.

[0214] Statement 19: The method of Statement 14, wherein the Theileria is Theileria annae, Theileria annulata, Theileria buffeli, Theileria equi, Theileria haneyi, Theileria lestoquardi, Theileria luwenshuni, Theileria mutans, Theileria orientalis, Theileria ovis, Theileria parva, Theileria separata, Theileria sergenti, Theileria taurotragi, Theileria uilenbergi, or Theileria velifera.

[0215] Statement 20: The method of Statement 14, wherein the Trypanosome is Trypanosoma brucei, Trypanosoma congolense, Trypanosoma cruzi, Trypanosoma evansi, Trypanosoma equiperdum, Trypanosoma rangeli, Trypanosoma simiae, or Trypanosoma vivax.

[0216] Statement 21: The method of Statement 10, wherein the arthropod-borne pathogen is a virus and the virus is African horse sickness virus, African swine fever virus, Aino virus (AINV), Akabane virus (AKAV), Alkhurma hemorrhagic fever virus, Barmah forest virus (BFV), Bebaru virus (BENV), Bhanja virus, Bluetongue virus, Bourbon virus, Bovine ephemeral fever virus, Bunyamwera virus (BUNV), Cache Valley virus (CVV), Chandipura virus (CHPV), Chikungunya virus (CHIKV), Colorado tick fever virus, Crimean Congo hemorrhagic fever virus (CCHFV), Dengue virus (DENV), Eastern equine encephalitis virus (EEEV), Edge Hill virus, Equine encephelitis virus, Everglades virus (EVEV), Getah virus (GETV), Guaroa virus (GROV), Heartland virus, Ibaraki virus, Ilheus virus (ILHV), Jamestown Canyon virus (JCV), Japanese encephalitis virus (JEV), Kairi virus, Kasba virus, Kyasunur Forest disease virus (KFDV), La Crosse virus (LACV), Main drain virus (MDV), Mayaro virus (MAYV), Middelburg virus (MIDV), Murray Valley encephalitis virus (MVEV), Nairobi sheep disease virus, Ndumu virus (NDUV), Omsk hemorrhagic fever virus, O'nyong nyong virus (ONNV), Oropouche virus (OROV), Peruvian horse sickness virus, Powassan virus (POWV), Rift Valley fever virus (RVFV), Ross River virus (RRV), Sandfly fever virus, Schmallenberg virus (SB V), Semliki forest virus (SFV), Shuni virus, Sindbis virus (SINV), Snowshoe hare virus (SSHV), St. Louis encephalitis virus (SLEV), Tembusu virus, Thogoto virus, Tick borne encephalitis virus (TBEV), Tyuleniy virus, UNA virus (UNAV), Usutu virus (USUV), Venezualan equine encephalitis virus (VEEV), Vesicular stomatitis virus (VSV), Wesselsbron virus, West Nile virus (WNE), Westernequine encephalitis virus (WEEV), Yellow fever virus (YFV), Yunnan virus, or Zika virus (zncv).

[0217] Statement 22: The method of any of Statements 1-21, wherein the PCR primers specific for at least three arthropod species hybridize with a gene from a ribosomal 12S subunit, a ribosomal 16S subunit, a ribosomal 18S subunit, or a cytochrome c oxidase unit 1 of the at least three arthropod species.

[0218] Statement 23: The method of Statement 2, wherein the tick species comprise a soft tick or a hard tick.

[0219] Statement 24: The method of Statement 23, wherein the tick species is Amblyomma americanum, Amblyomma cajennense, Amblyomma maculatum, Amblyomma sp., Amblyomma testudinarium, Amblyomma variegatum, Argas persicus, Bothriocroton hydrosauri, Dermacentor albipictus, Dermacentor andersoni, Dermacentor nitens, Dermacentor occidentalis, Dermacentor reticulatus, Dermacentor sp., Dermacentor variabilis, Haemaphysalis longicornis, Haemaphysalis sp., Hyalomma marinatum, Hyalomma sp., Hyalomma truncation, Ixodes cookei, Ixodes holocyclus, Ixodes pacificus, Ixodes persulcatus, Ixodes ricinus, Ixodes scapularis, Ixodes sp., Ixodes spinipalpis, Margaropus winthemi, Ornithodoros moubata, Ornithodoros rudis, Ornithodoros savignyi, Ornithodoros turicatae, Otobius megnini, Rhipicephalus annlatus, Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Rhipicephalus sanguineus, or Rhipicephalus sp.

[0220] Statement 25: The method of Statement 3, wherein the mosquito is an Aedes, an Anopheles, a Coquillettidia, a Culex, a Culiseta, a Haemagogus, an Ochlerotatus albifasciatus, or a Psorophora mosquito.

[0221] Statement 26: The method of Statement 25, wherein the Aedes, the Anopheles, the Coquillettidia, the Culex, the Culiseta, the Haemagogus, the Ochlerotatus, or the Psorophora mosquito is Aedes aegypti, Aedes albopictus, Aedes bahamensis, Aedes fulvus pallens, Aedes infirmatus, Aedes sollicitans, Aedes sp., Aedes tormentor, Aedes taeniorhynchus, Aedes triseriatus, Aedes vexans, Anopheles albimanus, Anopheles arabiensis, Anopheles atroparvus, Anopheles barbirostris, Anopheles christyi, Anopheles claviger, Anopheles coluzzii, Anopheles coustani, Anopheles culicifacies, Anopheles darlingi, Anopheles dims, Anopheles epiroticus, Anopheles farauti, Anopheles funestus, Anopheles gambiae, Anopheles maculatus, Anopheles melas, Anopheles merus, Anopheles minimus, Anopheles nimbus, Anopheles pseudopunctipennis, Anopheles quadriannulatus, Anopheles rufipes, Anopheles sacharovi, Anopheles sinensis, Anopheles sp., Anopheles stephensi, Anopheles walker, Coquillettidia perturbans, Coquillettidia sp., Coquillettidia venezuelensis, Culex biscaynensis, Culex declarator, Culex erraticus, Culexfatigans, Culexpedroi, Culex pipiens, Culex quinquefasciatus, Culex sp., Culiseta dyari, Culiseta inornata, Culiseta melanora, Culiseta sp., Haemagogus capricornii, Haemagogus equinus, Haemagogus janthinomys, Haemagogus lucifer, Haemagogus sp., Oehler otatus albifasciatus, Oehler otatus sp., Ochlerotatus triser iatus, Psorophora confmnis, Psorophora ferox, or Psorophora sp.

[0222] Statement 27: The method of Statement 4, wherein the biting midge, the sandfly, or the tsetse fly are Culicoides sp., Glossina sp., Lutzomyia sp. or Phlebotomus sp.

[0223] Statement 28: The method of Statement 27, wherein the Culicoides sp. biting midge is Culicoides actoni, Culicoides adersi, Culicoides brevitarsis, Culicoides fulvus, Culicoides furans, Culicoides grahamii, Culicoides imicola, Culicoides inornatipennis, Culicoides insignis, Culicoides insinuatus, Culicoides milnei, Culicoides obsoletus, Culicoides oxystoma, Culicoides paraensis, Culicoides phlebotomus, Culicoides schultzei, Culicoides variipennis, or Culicoides wadai.

[0224] Statement 29: The method of Statement 27, wherein the Glossina sp. tsetse fly is Glossina austeni, Glossina brevipalpis, Glossina fusca, Glossina fuscipes, Glossina longipalpis, Glossina morsitans, Glossina palpalis, Glossina tabaniformis, Glossina tachinoides, or Glossina vanhoofi.

[0225] Statement 30: The method of Statement 27, wherein the Lutzomyia sp. o Phlebotomus sp. sandfly is Lutzomyia amazonensis, Lutzomyia anduzei, Lutzomyia aracuchensis, Lutzomyia ayrozai, Lutzomyia carerrai, Lutzomyia christophei, Lutzomyia complexa, Lutzomyia diabolica, Lutzomyia evansi, Lutzomyia flaviscutellata, Lutzomyia gomezi, Lutzomyia hartmanni, Lutzomyia intermedia, Lutzomyia llanosmartinsi, Lutzomyia longipalpis, Lutzomyia migonei, Lutzomyia panamensis, Lutzomyia paraensis, Lutzomyia peruensis, Lutzomyia pessoai, Lutzomyia trapidoi, Lutzomyia umbratilis, Lutzomyia wellcomei, Lutzomyia whitmani, Lutzomyia whitmani, Lutzomyia ylephiletor, Lutzomyia ylephiletor, Lutzomyia yucumensis, Lutzomyia olmeca, Phlebotomus alexandri, Phlebotomus ansarii, Phlebotomus argentipes, Phlebotomus ariasi, Phlebotomus caucasicus, Phlebotomus celiae, Phlebotomus chinensis, Phlebotomus duboseqi, Phlebotomus kandelakii, Phlebotomus langeroni, Phlebotomus longicuspis, Phlebotomus longiductus, Phlebotomus longipes, Phlebotomus martini, Phlebotomus near rossi, Phlebotomus neglectus, Phlebotomus orientalis, Phlebotomus papatasi, Phlebotomus pedifer, Phlebotomus perfiliewi, Phlebotomus perniciosus, Phlebotomus salehi, Phlebotomus sergenti, Phlebotomus smirnovi, Phlebotomus tobbi, Phlebotomus transcaucasicus, or Phlebotomus vansomeranae.

[0226] Statement 31: The method of Statement 5, wherein the flea is Aetheca wagneri, Amphipsylla sp., Anomiopsyllus sp., Atyphloceras sp., Callopsylla sp., Catallagia sp.,Ceratophyllus sp., Chiastopsylla sp., Citellophilus sp., Coptopsylla sp., Craneopsylla sp., Ctenocephalides sp., Ctenophthalmus sp., Diamanus montanus, Dinopsyllus sp., Eumolpianus eumolpi, Foxella sp., Frontopsylla sp., Hectopsylla sp., Hoplopsyllus sp., Hystrichopsylla sp., Listropsylla sp., Malaraeus sp., Megabothris sp., Megarthroglossus sp., Meringis sp., Monopsyllus sp., Neopsylla sp., Neotyphloceras sp., Nosopsyllus sp., Odontopsyllus sp., Opisocroslis spp., Opisodasys sp., Orchopeas sp., Oropsylla sp., Parapsyllus sp., Pleochaetis sp., Polygenis sp., Psocopsylla sp., Pulex irritans, Rhadinopsylla sp., Stivalius sp., Synosternus sp., Thrassis sp., Tiamastus sp., Tritopsylla sp., Xenopsylla cheopis, or Xenopsylla sp.

[0227] Statement 32: The method of Statement 5, wherein the louse is a member of a Haematopinidae, Hoplopleuridae, Linognathidae, Pediculidae, Polyplacidae, or Pthiridae family.

[0228] Statement 33: The method of Statement 32, wherein the member of the Haematopinidae, Hoplopleuridae, Linognathidae, Pediculidae, Polyplacidae, Pthiridae family is Haematopinus asini, Haematopinus eurysternus, Haematopinus quadripertusus, Haematopinus suis, Haematopinus tuberculatus, Haematopinus tuberculatus, Haemodipsus ventricosus, Hoplopleura captiosa, Hoplopleura pacifica, Linognathus africanus, Linognathus africanus, Linognathus ovillus, Linognathus pedalis, Linognathus setosus, Linognathus stenopsis, Linognathus vituli, Pediculus humanus capitis, Pediculus humanus humanus, Polyplax serrata, Polyplax spinulosa, Pthirus pubis, Solenopotes capillatus

[0229] Statement 34: The method of Statement 5, wherein the triatomine bug is Panstrongylus megislus, Rhodnius prolixus, Triatoma brasiliensis, Triatoma dimidiate, or Triatoma infestans.

[0230] Statement 35: The method of any of Statements 1-34, wherein the PCR primers specific for at least three blood meal hosts species hybridize with a satellite DNA, a tandem repeat, or a mitochondrial gene from the blood meal host species.

[0231] Statement 36: The method of any of Statements 1-35, wherein the arthropod bloodmeal host species is an ape, a bat, a bear, a bird, a bovine, a camelid, a canid, a cervid, a dog, an equid, a felid, a fox, a giraffe, a goat, a horse, a human, a monkey, an opossum, a pig, a rabbit, a raccoon, a rodent, a sheep, a shrew, or a wild boar.

[0232] Statement 37: The method of Statement 36, wherein the bird is a chicken, a crane, a duck, an emu, a goose, a pheasant, a quail, or a turkey.

[0233] Statement 38: The method of Statement 36, wherein the bovine is an auroch, a bison, a buffalo, a domestic cow, an eland, a four-horned antelope, a guar, a spiral-horned antelope, a waterbuck, or a yak.

[0234] Statement 39: The method of Statement 36, wherein the camelid is an alpaca, a Bactrian camel, a dromedary, a guanaco, a llama, or a vicuna.

[0235] Statement 40: The method of Statement 36, wherein the canid is a coyote, a dog, a jackal, or a wolf.

[0236] Statement 41: The method of Statement 36, wherein the cervid is a brocket, a chital, an elk, a montjac, a moose, a mule deer, a red deer, a reindeer, a roe deer, a seka deer, or a whitetailed deer.

[0237] Statement 42: The method of Statement 36, wherein the equid is a donkey, a horse, a mule, or a zebra.

[0238] Statement 43: The method of Statement 36, wherein the felid is a bobcat, a cougar, a domestic cat, a jaguar, a leopard, a lion, a lynx, a mountain lion, an ocelot, a panther, a puma, or a tiger.

[0239] Statement 44: The method of Statement 36, wherein the rodent is a capybara, a chipmunk, a gerbil, a marmot, a mouse, a prairie dog, a rat, a squirrel, or a vole.

[0240] Statement 45: The method of Statement 44, wherein the mouse is a North American deer mouse or a white-footed mouse.

[0241] Statement 46: The method of any of Statements 1-45, wherein the extended products are analyzed by mass spectrometry.

[0242] Statement 47: The method of Statement 46, wherein the mass spectrometry is MALDI-TOF mass spectrometry.

[0243] Statement 48: The method of any of Statements 1-47, wherein the PCR primers are selected from nucleotide sequences set forth in SEQ ID Nos: 2, 3, 10, 11, 12, 13, 15, 16, 20, 21, 23, 24, 25, 26, 28, 29, 33, 34, 40, 41, 42, 44, 46, 47, 53, 54, 57, 60, 61, 62, 63, 69, 70, 74, 75, 81, 82, 84, 85, 86, 89, 90, 91, 92, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 107, 108, 109, 111, 112, 115, 116, 118, 119, 120, 122, 123, 126, 127, 128, 129, 131, 132, 134, 136, 137, 138, 139, 140, 141, 142, 144, 145, 146, 148, 149, 151, 152, 153, 154, 156, 157, 160, 161, 166, 167, 169, 170, 173, 174, 178, 179, 180, 181, 182, 183, 186, 187, 188, 189, 194, 195, 200, 201, 203, 204, 206, 207, 209, 210, 212, 213, 215, 216, 218, 219, 221, 222, 224, 225, 226, 228, 229, 231, 232, 236, 237, 239, 240, 242, 243, 244, 245, 248, 249, 252, 253, 256, 257, 260, 261, 263, 264, 265, 266, 269, 270, 273, 274, 277, 278, 280, 281, 282, 283, 286, 287, 289, 290, 293, 294, 298, 299, 302, 303, 305, 306, 307, 308, 310, 311, 315, 316, 320, 321, 324, 325, 328, 329, 332, 333, 336, 337, 339, 340, 341, 342, 346, 347, 348, 349, 351, 352, 355, 356, 357, 358, 359, 360, 361, 362, 366, 367, 368, 369, 373, 374, 375, 376, 378, 379, 381, 382, 385, 386, 388, 389, 390, 392, 393, 397, 398, 401, 402, 403, 404, 407, 408, 411, 412, 415, 416, 417, 418, 421, 422, 425, 426, 427, 428,429, 430, 433, 434, 436, 437, 438, 439, 443, 444, 445, 446, 450, 451, 452, 453, 454, 456, 457, 458, 462, 463, 464, 465, 468, 469, 472, 473, 476, 477, 480, 481, 484, 485, 487, 488, 490, 491, 494, 495, 498, 499, 501, 502, 504, 505, 507, 508, 509, 510, 511, 512, 514, 515, 518, 519, 520, 522, 523, 524, 525, 529, 530, 531, 532, 535, 536, 538, 539, 541, 542, 543, 544, 546, 547, 548, 549, 553, 554, 555, 556, 558, 559, 560, 561, 562, 563, 566, 567, 568, or 569.

[0244] Statement 49: The method of Statement 48, wherein the extension primers are selected from nucleotide sequences set forth in SEQ ID NOs:1, 8, 9, 14, 19, 22, 30, 31, 32, 39, 43, 45, 48, 49, 50, 52, 55, 58, 64, 65, 66, 67, 68, 71, 72, 76, 80, 83, 88, 93, 106, 110, 113, 114, 117, 121, 124, 125, 130, 133, 135, 143, 147, 150, 155, 158, 159, 165, 168, 172, 175, 177, 184, 185, 192, 193, 199, 202, 205, 208, 211, 214, 217, 220, 223, 227, 230, 235, 238, 241, 246, 247, 250, 251, 254, 259, 262, 268, 272, 276, 279, 285, 288, 292, 296, 297, 301, 304, 309, 314, 317, 319, 322, 323, 330, 331, 335, 338, 343, 344, 345, 350, 353, 354, 363, 364, 365, 370, 371, 372, 377, 380, 383, 391, 394, 396, 399, 405, 409, 413, 419, 423, 424, 431, 432, 435, 440, 441, 442, 447, 449, 455, 459, 461, 466, 471, 475, 479, 483, 486, 489, 493, 496, 497, 500, 503, 506, 513, 517, 521, 526, 527, 534, 537, 540, 545, 550, 552, 557, 564, or 565.

[0245] Statement 50: The method of any of Statements 1-49, wherein the amplified products or the extended products are analyzed by next generation sequencing.

[0246] Statement 51: The method of Statement 50, wherein the amplified products are selected from nucleotide sequences set forth in SEQ ID Nos: 4, 5, 6, 7, 17, 18, 27, 35, 36, 37, 38, 51, 56, 59, 73, 77, 78, 79, 87, 104, 105, 162, 163, 164, 171, 176, 190, 191, 196, 197, 198, 233, 234, 255, 258, 267, 271, 275, 284, 291, 295, 300, 312, 313, 318, 326, 327, 334, 384, 387, 395, 400, 406, 410, 414, 420, 448, 460, 467, 470, 474, 478, 482, 492, 516, 528, 533, or 551.

[0247] Statement 52: The method of any of Statements 1-51, wherein the biological sample is a biopsy, a blood sample, a cerebral spinal fluid (CSF) sample, an environmental sample, a fluid from a skin blister or ooze from a lesion, a milk sample, a saliva sample, a sewage sample, a synovial fluid sample, a tick homogenate, a tissue sample, or a urine sample.

[0248] Statement 53: The method of Statement 52, wherein the blood sample is a plasma sample, a peripheral blood mononuclear cell (PBMC) sample, a serum sample, or a whole blood sample.

[0249] Statement 54: The method of Statement 53, wherein the method is able to identify an arthropod-borne pathogen in a 1 mL blood sample if greater than 10 arthropod-borne pathogen genomes are present in the blood sample.

[0250] Statement 55: The method of any of Statements 1-54, wherein multiple arthropod-borne pathogens are present in the biological sample.

[0251] Statement 56: The method of any of Statements 1-55, wherein the biological sample is a pooled sample.

[0252] Statement 57: The method of any of Statements 1-55, wherein the biological sample is a sample from a single subject.

[0253] Statement 58: The method of any of Statements 1-57, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 24 hours.

[0254] Statement 59: The method of Statement 58, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 12 hours.

[0255] Statement 60: The method of Statement 58, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 8 hours.

[0256] Statement 61: A method for diagnosing an arthropod-related disease which comprises: (a) identifying the arthropod and the arthropod-borne pathogen in the biological sample from a human patient or an animal subject by the method of any of Statements 1-60; and (b) using the identification of the arthropod and the arthropod-borne pathogen to diagnose the arthropod-related disease.

[0257] Statement 62: The method of Statement 61, wherein the arthropod-related disease is African swine fever, Alkhurma hemorrhagic fever, anaplasmosis, babesiosis, bluetongue disease, chikungunya, dengue, ehrlichiosis, encephalitis, equine encephalitis, hepatozoonosis, Lyme disease, malaria, Oropouche fever, plague, rickettsiosis, Rocky Mountain Spotted Fever, tick-borne relapsing fever, tularemia, West Nile, yellow fever, or zika fever.

[0258] Statement 63: The method of any of Statements 60 or 61, wherein the human patient is a patient who (i) experienced an arthropod bite; (ii) is suspected of having been bitten by an arthropod; or (iii) is experiencing fever / chills, joint pain, muscle aches, persistent fever, or rash.

[0259] Statement 64: The method of any of Statements 60 or 61, wherein the animal subject (i) experienced an arthropod bite; (ii) is suspected of having been bitten by an arthropod; or (iii) is showing signs of lethargy, lameness, fever, joint pain or swelling, or swollen lymph nodes.

[0260] Statement 65: The method of any of Statements 60-64, wherein the arthropod-borne disease is an acute or a chronic an arthropod-borne disease.

[0261] Statement 66: A method of disease vector surveillance which comprises analyzing biological samples by the method of any of Statements 1-60 in order to identify an arthropod species or an animal host that may act as disease vectors.

[0262] Statement 67: A method of sentinel surveillance which comprises analyzing biological samples from a sentinel source by the method of any of Statements 1-60 in order to identify the arthropod species, the arthropod-borne pathogen(s), and / or the blood meal hosts in the sentinel source.

[0263] Statement 68: A method of monitoring an eradication program which comprises analyzing biological samples by the method of any of Statements 1-60 in order to identify the arthropod species, the arthropod-borne pathogen(s), and the blood meal hosts.

[0264] Statement 69: A kit for identification of an arthropod species, an arthropod-borne pathogens, or a blood meal hosts from a single biological sample, wherein the kit comprises: (a) a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three species of arthropods, and (iii) PCR primers specific for at least three host species; (b) a set of extension primers; and (c) appropriate reagents and instructions for a user to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).

[0265] It should be understood that the above description is only representative of illustrative embodiments and examples. For the convenience of the reader, the above description has focused on a limited number of representative examples of all possible embodiments, examples that teach the principles of the disclosure. The description has not attempted to exhaustively enumerate all possible variations or even combinations of those variations described. That alternate embodiments may not have been presented for a specific portion of the disclosure, or that further undescribed alternate embodiments may be available for a portion, is not to be considered a disclaimer of those alternate embodiments. One of ordinary skill will appreciate that many of those undescribed embodiments, involve differences in technology and materials rather than differences in the application of the principles of the disclosure. Accordingly, the disclosure is not intended to be limited to less than the scope set forth in the following claims and equivalents.

[0266] INCORPORATION BY REFERENCE

[0267] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entireties for all purposes. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world. It is to be understood that, while the disclosure has been described inconjunction with the detailed description, thereof, the foregoing description is intended to illustrate and not limit the scope. Other aspects, advantages, and modifications are within the scope of the claims set forth below. All publications, patents, and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

[0268] APPENDIX A SequencesSEQUENCE DATA

[0269] I hereby state that the information recorded in computer readable form is identical to the written sequence listing below.

[0270] Sequence nomenclature / abbreviations: amp = amplicon, ext = extension primer, F = PCR forward primer, R = PCR reverse primer. The underlined sequences in the amplicons are the forward and reverse primers. The bold sequences in the amplicons are the primer extension sequences. Bases in italics represent sequence variants. Table 1 below shows the SEQ ID NO. and the sequence name.ARTHROPOD-BORNE PATHOGENS SEQUENCES (SEQ ID NOS: 1-376) < Anaplasma phagocytophilum 16S ext 1; DNA; Anaplasma phagocytophilum> CGCCTTCGCCACTGGTGT TCCTC “ “ “< Anaplasma phagocytophilum 16S F 1; DNA; Anaplasma phagocytophilum> ACGTTGGATG CGGGAGAGGATAGCGGAATTC< Anaplasma phagocytophilum 16S R 1; DNA; Anaplasma phagocytophilum> ACGTTGGATG CGTCAGTACCGGACCAGATAG< Anaplasma phagocytophilum amp 1; DNA; Anaplasma phagocytophilum> CCAGCGTTTAGCAAGATAAGAGATTTTAGTATAAGGGAGAGTAACGGAGAGACTAAGGCAGTATATCCAT ACTTAAAGGATGGAAAGAGTGTAAAGCTAGAGTCACACAAGTTTGACTGGAACACTCCTGATCCTCGGAT TGGGTTTAAGGACAACATGCTTGTAGCTATGGAAGGCAGTGTTGGTTATGGTATTGGTGGTGCCAGGGTT GAGCTTGAGATTGGTTACGAGCGCTTCAAGACCAAGGGTATTAGAGATAGTGGTAGTAAGGAAGATGAAG CTGATACAGTATATCTACTAGCTAAGGAGTTAGCTTATGATGTTGTTACTGGGC< Anaplasma phagocytophilum amp 2; DNA; Anaplasma phagocytophilum> GGTAAAGCATGTAAAGTACTACTAAAGGCTGGGGCGTCAGTATCAGTCGTGAATGTAGAGGGAAAAACAC CGGTAGATGTTGCAGATCCATCATTGAAAGCTCGTCCGTGGCTTTTTGGAAAGTCCGTTGTCACAATGAT GGCTGAACGTGTTCAAGTTCCTGAAGGGGGATTTCCTCCATATCTGCCGCCTGAAAGTCCAACTCCTTCT TTAGGATCTATTTCAAGTTTTGATAGTGTCTCTGTGCTATCATCCTTGGGTAGTGGCCTAGACACTGCAG< Anaplasma phagocytophilum amp 3; DNA; Anaplasma phagocytophilum> GTTCGGTAAGTTAAAGGTGAAATGCCAGGGCTTAACCCTGGAGCTGCTTTTAATACTGCCAGACTAGAGA CCGGGAGAGGATAGCGGAATTCCTAGTGTAGAGGTGAAATTCGTAGATATTAGGAGGAACACCAGTGGCG AAGGCGGCTATCTGGTCCGGTACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCC TGGTAGTCCACGCTGTAAACGATGAGTGCTGAATGTGGGGATATTTTATCTCTGTGTTGAAGCTAACGCG< Anaplasma phagocytophilum amp 4; DNA; Anaplasma phagocytophilum> CTTTGGGCTTTCGTCGCACAAACCTTCTTATCAATACCGGGATGAGAAACCACAACCGCCTTAGCAAACT GAACAAAGTCTTTACCGGAAGTTTTGGCAAGAGCAGCAGCAAGGTTATCAGTCTGTCCAGTAACAACATC ATAAGCTAACTCCTTAGCTAGTAGATATACTGTATCAGCTTCATCTTCCTTACTACCACTATCTCTAATA CCCTTGGTCTTGAAGCGCTCGTAACCAATCTCAAGCTCAACCCTGGCACCACCAATACCATAACCAACAC< Anaplasma phagocytophilum msp2 ext 1; DNA; Anaplasma phagocytophilum> TTATCAGTCTGTCCAGTAAC - - - < Anaplasma phagocytophilum msp2 ext 2; DNA; Anaplasma phagocytophilum> TGGTGCCAGGGTTGAGCTTGA< Anaplasma phagocytophilum msp2 F 1; DNA; Anaplasma phagocytophilum> ACGTTGGATG ATGGAAGGTAGTGTTGGTTATGGTATT< Anaplasma phagocytophilum msp2 F 2; DNA; Anaplasma phagocytophilum> AC GT T G GAT G T G GAC AT T GAGAGT AC AT GT G G GA< Anaplasma phagocytophilum msp2 R 1; DNA; Anaplasma phagocytophilum> ACGTTGGATG TTGGTCTT GAAGCGCTCGTA”< Anaplasma phagocytophilum msp2 R 2; DNA; Anaplasma phagocytophilum> ACGTTGGATG CCTCTTGCTGTCCTATCTCTTGCT< Babesia microti 18S ext 1; DNA; Piroplasmida>ACACCGCCCGTCGCTCCTA< Babesia microti 18S F 1; DNA; Piroplasmida>ACGTTGGATG CGACTACGTCCCTGCCCTTTG< Babesia microti 18S R 1; DNA; Piroplasmida>ACGTTGGATG ACGAAGGACGAATCCACGTTTC< Babesia microti amp 1; DNA; Piroplasmida> AACGAGGAATGCCTAGTAGGCGCGAGTCATCAGCTCGTGCCGACTACGTCCCTGCCCTTTGTACACACCG CCCGTCGCTCCTACCGATCGAGTGATCCGGTGAATTATTCGGACCAAGAAACGTGGATTCGTCCTTCGTT TTTTGGAAAGTTTTGTGAACCTTATCACTTAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTGAAC CTGCGGAAGGATCATTCTTATCAGAGTTCTTTGTATCCCATTTGGGTTACGCTGGCCCGTGCGCTTGGTC< Babesia microti amp 2; DNA; Piroplasmida> CCTTACATGGCGTTATCATGATATTCTTCAACATCATGCCAGGCCTGTTTGGAGGTATAGGTAACTACCT ACTACCCATACTGGTCGGTGCTCCCGAGGTTGTCTTCCCAAGAGCTAACTTGTATAGTCTGTTGTTACAA C CACTAGCATTTGCTCTTGTGGTTTCTTCACTCTACTGT GAAGTAAGT GGAGGTACAGGAT GGACATTAT< Babesia microti coxl ext 1; DNA; Piroplasmida> TACTACCCATACTGGTCGGTGCT< Babesia microti coxl F 1; DNA; Piroplasmida>ACGTTGGATG CATCATGCCAGGCCTGTTTG< Babesia microti coxl R 1; DNA; Piroplasmida>ACGTTGGATG GAAGAAACCACAAGAGCAAATGC< Babesia spp ext 1; DNA; Piroplasmida>GCTTCTTAGAGGGAC- < Babesia spp 18S F 1; DNA; Piroplasmida>CAT GAACGAGGAAT GCCTAGTAT G< Babesia spp 18S R 1; DNA; Piroplasmida>CCGAATAAT TCA CCG GAT CAC TC< Babesia spp F 1; DNA; Piroplasmida>ACGTTGGATG ACGAGACCT / GTAACCTGCTAA< Babesia spp F 1; DNA; Piroplasmida>ACGTTGGATG CACAGACCTGTTATTGCCT / GTA< Borrelia afzelii amp 1; DNA; Borrelia afzelii> TTATAAATTCTTGTGGTCCTGGTTCTTTTACTGGTCTTAGAGTTAGTTTGAGTTTTGTAAAAGGCCTTGC TTTAGGTCTTTCTATTCCTTTTGTCAATATTTCTACATTGGATGTTTTTGCAAATTTATTTAAAGATAGT TCTAGTGTAATTGTATTAACTTTTACTGCTGGCAAATATTTTCTTGGGCATTATAAAAATCTCGAATTGG T AGGT AAGAT TTTGTGTTTT T CT AAGGAAGAT T T GT T T GAATATCTAGGGCAGATTGATTCAAAT T CAGT GCTTACTGGTTACAATCTTGAGAATGTTTGTAAGGAATTTAATTATAAATTTAAAATCATTGAAAATTTG< Borrelia afzelii F 1; DNA; Borrelia afzelii>ATTCTTGTGGTCCTGGTT- < Borrelia afzelii R 1; DNA; Borrelia afzelii>CTAGGGCAGATTGATTCA< Borrelia afzelii tsaB ext 1; DNA; Borrelia afzelii> CTGGTCTTAGAGTTA “ “ “< Borrelia americanam IST1 ext 1; DNA; Borrelia americanam> gggGTTAGTGTTTTTAAATGTG< Borrelia burgdorferi 23S ext 1; DNA; Borrelia burgdorferi> AGATGTGGTAGACCCGAAG “ “ “< Borrelia burgdorferi 23S F 1; DNA; Borrelia burgdorferi> ACGTTGGATG CGAGTCTTAAAAGGGCGATTTAGT< Borrelia burgdorferi 23S R 1; DNA; Borrelia burgdorferi> ACGTTGGATG GCTTC AGCCTGGCCATAAATAG< Borrelia burgdorferi amp 1; DNA; Borrelia burgdorferi> GTACCCTTTGACAGGCCCTAGCTCAATTAGTGCTCTACCTCTATTAAACTAAAATAAGGCTGAACTTAAA TCCATTTCGGGGAGAACCAGCTATCTCCGAGTTTGTTTAGCCTTTCACTCCTATTCACAGCTCATCCCTG CCTTTTTAAACAGACTAGAGTTCGGCCCTCCACTTGGTTTTACCCAAGCTTCAGCCTGGCCATAAATAGA TCACTCGGCTTCGGGTCTACCACATCTAACTAAATCGCCCTTTTAAGACTCGCTTTCGCTTCGACTCCAG CACTTCTATGCTTTAATCTTGCTAGACATGATAACTCGCAGGTTCATTATGCAAAAGGCACGCCATCACC< Borrelia burgdorferi amp 2; DNA; Borrelia burgdorferi>AAAT GAAC GT T CT T GTAAGCAAAGAAAAAAACAAAGACGGCAAGTACGATCTAAT T GCAACAGTAGACAA GCTTGAGCTTAAAGGAACTTCTGATAAAAACAATGGATCTGGAGTACTTGAAGGCGTAAAAGCTGACAAA AGTAAAGTAAAATTAACAATTTCTGACGATCTAGGTCAAACCACACTTGAAGTTTTCAAAGAAGATGGCA< Borrelia burgdorferi amp 3; DNA; Borrelia burgdorferi> CTAGAATTTTATCTAAGCAATGACAAAACATATTGGGGAACTTGATTAGCCTGCGCAATCATTGCCATTG CAGATTGTGTTAAAATACTATTAGTTGTTGCTGCTACAACCTCATCTGTCATTGTAGCATCTTTTATTTG AGCATAAGATGCTTTTAGATTTTCAATTGCATACTCAGTACTATCCTTTATAGATTCAAGTCTATTTTGG AAAGCACCTAAATTTGCTCTTTGATCACTTATCATTCTAATAGCATTTTCAATTTTAGCAAGTGATGTAT< Borrelia burgdorferi amp 4; DNA; Borrelia burgdorferi>AGCT CAACAGCGTATAAAAT GTAT GAAAAT GAAGAGCTAGATGCAATTTTTGGTTCCATACCCCCAGAT C TAATCAAAAATCTAAAATTAAGAAGCGACTATTACTCATCAGCTGTTAATGCCATATACTTTTACGCGTT C AATAC AC AC ATC AAAC C AC T T GAC AAC GT T AAAAT T AGAAAAG C CTTAACTCTTGCTATTGACAGAGAA< Borrelia burgdorferi flagrin ext 1; DNA; Borrelia burgdorferi> C AAC CTC ATC TGT CAT TGT AG~< Borrelia burgdorferi flagrin F 1; DNA; Borrelia burgdorferi> ACGTTGGATG AGC AAA TTT AGG TGC TTT CCA A< Borrelia burgdorferi flagrin R 1; DNA; Borrelia burgdorferi> ACGTTGGATG GCA ATC ATT GCC ATT~ GCA GA< Borrelia burgdorferi mayonii R 1; DNA; Borrelia burgdorferi> ACGTTGGATG CTG TCA ATA GCA AGA~ GTT AA< Borrelia burgdorferi opp2 ext 1; DNA; Borrelia burgdorferi>CGT TCA ATA CAC ACA TCA AAC C~< Borrelia burgdorferi opp2 F 1; DNA; Borrelia burgdorferi> ACGTTGGATG AAT TTT TGG TTC CAT ACC C< Borrelia burgdorferi ospA ext 1; DNA; Borrelia burgdorferi>g g c C AAC AGT AGAC AAG C< Borrelia burgdorferi OspA F 1; DNA; Borrelia burgdorferi> ACGTTGGATG CCTTCAAGTACTCCAGATCCATTG< Borrelia burgdorferi OspA R 1; DNA; Borrelia burgdorferi> ACGTTGGATG AACAAAGACGGCAAGTACGATC< Borrelia californiensis IST1 1 ext 1; DNA; Borrelia sp. > ggTGTTAGTGTCTTTG< Borrelia carolinesis bissetti IST1 ext 1; DNA; Borrelia sp. > cccATTTTAAATCTTGA - - - < Borrelia carolinesis IST1 ext 2; DNA; Borrelia sp. > gcGTGTTTTTGAATATG< Borrelia garinii amp 1; DNA; Borrelia garinii>GAAT AGACAGAAGAAT AAAAAGT GAT AGAGAGT T C CAAAT T T T T GAAGGTAAAAAGATTAAGT T AAT G C T AGACAAT GAAT T T GAAGAG G GT T T T AT AT T AGAAT C T AAAT C AAAAAGT T T T AT T T T T AAAAC AGAAAG C AAAGAATTAAATGTTTTTTATAGCGATGTTAAAAAGGCTAGATTAGTTTAAGGAGGAAGTTTGGATGATA< Borrelia garinii rimP ext 1; DNA; Borrelia garinii> GAAGGTAAAAAGATTAAG “ “< Borrelia garinii rimP F 1; DNA; Borrelia garinii>ACGTTGGATG AAAAAGTGATAGAGAGTTCC< Borrelia garinii rimP R 1; DNA; Borrelia garinii>ACGTTGGATG CCCTCTTCAAATTCATTGTC< Borrelia hermsii ext 1; DNA; Borrelia hermsii>TGT TGA GAG TCT~ TAG~ AGC C< Borrelia hermsii amp 1; DNA; Borrelia hermsii> ATGGTATTGAACAGGTAAAAATATTTTTCCTTAATAGGGCAGATACTTTTATTAGAATAAAAGCAGGCGA TGTTCATTCCAGTGCTGACTTTTATTTAATTAATACTTTAATTTATAAAGATATTAAATTGCCCTTTAAA ATTAGTGATATTGCTACTGGTTCT T T T C T AGAAAT AG C T AGAT AT AT T AGT GAT T T T AT T GAT T T T GAG C TTTTTAATCCTAAATATCTTGAAGCTTATGATAATATTTATAATATTGTTGAGAGTCTTAGAGCCTTTTT AAAGTCTTCTCCTTTAATGATTGAGATGCATGATGGTGCAATGAATGAACTGGGTGAGATGGTATATATT AAGACAGGTGAGCCTCAAAGAGAACCTGTAGGATTTAATTGTTCTGGATTTGGCAAATGGGTAGCAGATT< Borrelia hermsii F 1; DNA; Borrelia hermsii>ACGTTGGATG ACGTTGGATG TTAGTGATATTGCTACTGGTTC< Borrelia hermsii flaB ext 1; DNA; Borrelia hermsii> CCTTCACCTGCAAAAAGA< Borrelia hermsii flaB amp 1; DNA; Borrelia hermsii> ATTAACTCCACCTTGAGCTGGAGCTGCTGCTGGAGCTGGAGCTGCTTGACCTTCCTCTTGCTGTCCTATC TCTTGCACTGGAGCAGCCTGAGCGCCTTCACCTGCAAAAAGATTTGCAACATTAGATGCATAAATATTAA CAGCAATTGCCTCATCCTGATTTGCGCCCACATGTACTCTCAATGTCCATGAAGCTTGTGATCCAGCTAG TGATGCTGGTGTGTTAATTTTTGCGGGTTGCATTCCAAGCTCTTCAGCTGTTTTTACATTTTGAGCAGCT< Borrelia hermsii flaB F 1; DNA; Borrelia hermsii>ACGTT GGAT G GGACATT GAGAGTACAT GT GGGC< Borrelia hermsii flaB R 2; DNA; Borrelia hermsii>ACGTTGGATG CCTCTTGCTGTCCTATCTCTTGCA< Borrelia hermsii parkeri F; DNA; Borrelia hermsii>ACGTTGGATG ACGTTGGATG TTAGTGATATTGCTACTGGTTC< Borrelia hermsii R 1; DNA; Borrelia hermsii>AT GAT GGT GCAAT GAAT GAACT< Borrelia kurtenbachii IST1 ext 1; DNA; Borrelia sp. > cccAAATGTTAGTGG - - - < Borrelia lanei IST1. 1 ext 1; DNA; Borrelia sp. >tccggTAATTTATGTTGC< Borrelia lanei IST1.2 ext 1; DNA; Borrelia sp. >cccggTAATTTATGTTGT< Borrelia lonestari fla ext 1; DNA; Borrelia sp. > cATCAACCAGCTCCAGCT “ “ “< Borrelia lonestari glpQ ext 1; DNA; Borrelia sp. > GAGCTAGGGAAGACGGACGATATT< Borrelia lonestari glpQ F 1; DNA; Borrelia sp. > GATCCAGAACTTGATACAACCACAA< Borrelia lonestari glpQ R 1; DNA; Borrelia sp. >T GAT T T AAGT T CAT CT AGT GT GAAGT CAGT< Borrelia lusitaniae IST1 ext 1; DNA; Borrelia sp. >gcccTCA AGA TTT GAA G ~ ~< Borrelia mayonii oppA2 ext 1; DNA; Borrelia sp. >CA CGC ACA TTA AAC CGC~ TTG~< Borrelia mayonii oppA2 amp 1; DNA; Borrelia sp. >G C AT AT AAAAT GT AT GAAAAT GAAGAAC T AGAC G C AAT C T T T G GAT C CAT AC CGCCCGATTTAATCAAAG ATCTAAAATTAAGAAGCGACTATTACTCATCAGCTGTTAATGCCATATACTTTTACGCATTTAACACGCA CATTAAACCGCTTGATAACGTTAAAATTAGAAAAGCCTTAACTCTTGCTATTGACAGAGAAACACTTACA< Borrelia mayonii oppA2 F 1; DNA; Borrelia sp. >ACGTTGGATG GCC CGA TTT AAT CAA AGA< Borrelia mayonii oppA2 R 1; DNA; Borrelia sp. > TTAACTCTTGCTATTGACAG< Borrelia miyamotoi ext 1; DNA; Borrelia miyamotoi> CACCTTAAAGTTCATGAG< Borrelia miyamotoi amp 1; DNA; Borrelia miyamotoi>AAAACCTTGAGGCTCTCTTTGAGGTTCACCAGTTTTAATATAGACCATCTCACCTAATTCATTCATAGCC< Borrelia miyamotoi amp 2; DNA; Borrelia miyamotoi>GCAACCT GCAAAAATTAACACACCAGCAT GATT GGCT GGAT CACAAGCTT CAT GGACATT GAGAGT GCAT GTAGGTGCAAATCAGGATGAAGCAATTGCTGTCAATATTTATGCAGCTAATGTTGCAAATCTTTTTAATG GAGAAGGTGCTCAAGCAGCTCCAGCTCAAGAGGGAGCACAACAGGAGGGAGTTCAAGCAGTTCCAGCTCC< Borrelia miyamotoi amp 3; DNA; Borrelia miyamotoi> GTTTTCAGGATCAAATCTTTCACTGAGACTTAGTGATTTAAGTTCAGTTAGTGTGAAGTCAGTGGCGTAA TATCGTCCGTTTTCTCTAGCTCGATTGGGAAATAATTGTGCAACATTTGTGGTTGTGTCAATTTCTGGGT CGTGCATTATAACAGGAATATTGTCCTTTGTTAGAACTATGTCTTGTTCTAGATAATCAGCTCCTAATGC< Borrelia miyamotoi flaB ext 1; DNA; Borrelia miyamotoi> TGTGGGTGCAAATCAGGATGAAG- < Borrelia miyamotoi flaB F 1; DNA; Borrelia miyamotoi>ACGTTGGATG AGCACAAGCTTCATGGACATTGA< Borrelia miyamotoi flaB R 1; DNA; Borrelia miyamotoi>ACGTTGGATG GAGCTGCTTGAGCACCTTCTC< Borrelia miyamotoi glpQ ext 1; DNA; Borrelia miyamotoi>CAA TCG AGC TAG AGA AAA~ CGG~< Borrelia miyamotoi glpQ F 1; DNA; Borrelia miyamotoi>ACGTTGGATG GAC CCA~ GAA ATT GAC ACA ACC ACA A< Borrelia miyamotoi glpQ R 1; DNA; Borrelia miyamotoi>ACGTTGGATG TGA TTT~ AAG TTC AGT TAG TGT GAA GTC AGT< Borrelia miyamotoi R 1; DNA; Borrelia miyamotoi>ACGTTGGATG ACGTTGGATG TCATTCATAGCCCCATCAT< Borrelia parkeri amp 1; DNA; Borrelia parkeri>AT AAAGAT AT T AAAT T G C C T T T TAAAATTAGTGATATTGCTACTGGTTCT T T T T T AGAAAT AG C T AAAT A TATTAGTAATTTTATTGATTTTGAACTTTTTAGACCCAGATCTCTTGAAGCTTATGATAATATTTCTAAT ATTGTTGACAGTCTGAGATCCTTTTTAAAGGTTTCTCCTTTAATATTTGAAGTACATGATGGAGCAATGA ATGAACTAGGTGAAATGGTGTATATTAGAACGGGTGAACCTCAAAGAGAACCTATAGGATTTAATTGTTC< Borrelia parkeri ext 1; DNA; Borrelia parkeri>AGA CC CAG ATC TCT TGA AG< Borrelia parkeri F 1; DNA; Borrelia parkeri>TTGTGATATTGCTATGGTTC< Borrelia parkeri R 1; DNA; Borrelia parkeri>AT GAT GGAGCAAT GA AT GA< Borrelia spp 0 F 1; DNA; Borrelia sp. >ACGTTGGATG TTG GTG ATA TTG CTA TTG GTT C< Borrelia spp 0 R 1; DNA; Borrelia sp. >ACGTTGGATG TCA TTC ATT GCA CCA TCA T< Borrelia spp 16 ext 1; DNA; Borrelia sp. >CG GCC TGA GAG GGT G< Borrelia spp 16S F 1; DNA; Borrelia sp. >GGC AAC CCT AAG GTG AAG GC< Borrelia spp 16S F 2; DNA; Borrelia sp. >ACGTTGGATG AGC YTT TAA AGC TTC GCT TGT AG< Borrelia spp 16S R 1; DNA; Borrelia sp. >GGT GAG CCA GGC CAT CAC TA< Borrelia spp 16S R 2; DNA; Borrelia sp. >ACGTTGGATG GCC TCC~CGT AGG AGT CTG G< Borrelia spp 1ST F 1; DNA; Borrelia sp. >ACGTTGGATG GAGTAAGTTATTGCCAGGG< Borrelia spp 1ST F 2; DNA; Borrelia sp. >ACGTTGGATG GAGTAGGTTATTGCCAGGG< Borrelia spp 1ST R 1; DNA; Borrelia sp. >ACGTTGGATG TCC ATC TCT ATT TTG CCA< Borrelia spp 1ST R 2; DNA; Borrelia sp. >ACGTTGGATG TCC ATC^CT ACT TTG CCA< Borrelia theileri lonestari F 1; DNA; Borrelia theileri> ACGTTGGATG AGCTCAAGAGGGTGCACAACAG< Borrelia theileri lonestari R 1; DNA; Borrelia theileri> ACGTTGGATG CGT M G~T / CGA GT / CT ATT AGC ATC< Borrelia turicatae amp 1; DNA; Borrelia turicatae>TTCGACTTTGACGAATTAAAGAGAATAAGGGAAGAACTTGGATATCAAGGAAAATTAATAATGCTTGTTGG C GAAAAT GAG T GGGATGAAGCACCAACAGACTAT GAAT AC AT AAAAT C AC AAGAAGGT TGAC GAGGT T G C AAAAT AT G C C GATGGAATTGGACCTTGGATACCCCAAAT T AT AAT T GAT G GAAAAAT AAC AGAT C T A< Borrelia turicatae amp 2; DNA; Borrelia turicatae> GTTATGAAGGAGAAATGACTACTAATACTCAAGATAGAACTTTTGTTGAGACCGGCACACAGGATTCTAA AACACAATAT T CT GAT T T T T CT GAT CAAGAT AT AAGAGAT AAAGT T T TGGGAAGTGTTGTTGGTGGC GT T GTAGACAATGTTATGAGTGGAATTGATAATGTTATTCAGGGAGCAGGAACTTTTGCTACAGCTGCTATGC< Borrelia turicatae bipA ext 1; DNA; Borrelia turicatae> TGGGAAGTGTTGTTGGTGGC< Borrelia turicatae bipA F 1; DNA; Borrelia turicatae>ACGTTGGATG AGACCGGTACACAGGATTCTAAAGC< Borrelia turicatae bipA F 2; DNA; Borrelia turicatae>ACGTTGGATG ACGTTGGATG CCGGCACACAGGATTCTAAAAC< Borrelia turicatae bipA R 1; DNA; Borrelia turicatae>ACGTTGGATG GTTCCTGCTCCCTGAATAACATTATC< Borrelia turicatae glpQ ext 1; DNA; Borrelia turicatae> GAAGGTATGACAGAGG - - - < Borrelia turicatae glpQ F 1; DNA; Borrelia turicatae>ACGTTGGATG GGATGAAGCACCAACAGACTA< Borrelia turicatae glpQ R 1; DNA; Borrelia turicatae>ACGTTGGATG GGGTATCCAAGGTCCAATTCC< Borrelia valaisiana IST1 ext 1; DNA; Borrelia valaisiana>cAA GGT TTG AAG TAC~< Bourbon virus 0 ext 1; DNA; Bourbon virus>CAGTTCAGCACCATAGATGCTG< Bourbon virus 0 F 1; DNA; Bourbon virus>ACGTTGGATG GATCCATTGCACCAACCCTTG< Bourbon virus 0 R 1; DNA; Bourbon virus>ACGTTGGATG TTGGTGATCTTCTCTACAGCC< Bourbon virus NP ext 1; DNA; Bourbon virus>TCACACCACGGAAGCTG< Bourbon virus NP F 1; DNA; Bourbon virus>ACGTTGGATG GCAAGAAGAGGCCAGATTTC< Bourbon virus NP R 1; DNA; Bourbon virus>ACGTTGGATG TCGAATTCAGCATTCAGAGC< Bourbon virus NP R 2; DNA; Bourbon virus>ACGTTGGATG TCGAATTCGGCATTCAGAGC< Bourbon virus PB1 ext 1; DNA; Bourbon virus> ACCCTTGCTGCATCTTCCACC- < Bourbon virus PB1 F 1; DNA; Bourbon virus>ACGTTGGATG AACCGAAGGACCATTGCTAC< Bourbon virus PB1 R 1; DNA; Bourbon virus>ACGTTGGATG ACAGGGACTCCAGAACTTGG< CTF virus ext 1; DNA; Colorado tick fever coltivirus> GATAGCTTCCCGTGGATATG< CTF virus ext 2; DNA; Colorado tick fever coltivirus> TGAAACTGCACGTACTCGAGCGGA< CTF virus F 1; DNA; Colorado tick fever coltivirus>ACGTTGGATG CTTGCTTCTTCCCGGATCAGT< CTF virus F 2; DNA; Colorado tick fever coltivirus>ACGTTGGATG TGACTGGGAATGTGAACTACGTGTAT< CTF virus R 1; DNA; Colorado tick fever coltivirus>ACGTTGGATG GTCGATTCGGTTTCCGGTAA< CTF virus R 2; DNA; Colorado tick fever coltivirus>ACGTTGGATG TCCCAACGGACTTGGACATC< Cytauxzoon felis 18S ext 1; DNA; Piroplasmida> Cgttgtggctttttctggtga< Cytauxzoon felis 18S F 1; DNA; Piroplasmida>ACGTTGGATG TGCATCATTTATATTCCTTAATCG< Cytauxzoon felis 18S R 1; DNA; Piroplasmida>ACGTTGGATG CAA TCT GGA~ TAA TCA TAG CGA A< DENV 1 ext 1; DNA; Orthoflavi virus denguei>cgTGTTTCCTGTTCCACAT< DENV 1 F 1; DNA; Orthoflavi virus denguei>ACGTTGGATG GYTTCCTGTGAGCCATTGTGAA< DENV all ext 1; DNA; Orthoflavi virus denguei>AGCATCATTCCAGGCA< DENV all F 1; DNA; Orthoflavi virus denguei>ACGTTGGATG” GARAGACCAGAGATCCTGCTGTCT< DENV all R 1; DNA; Orthoflavi virus denguei>ACGTTGGATG- ACCATTCCATTTTCTGGCGTT< DENV-13 R; DNA; Orthoflavi virus denguei>ACGTTGGATG GAATCTCTTCGCCAACTGTGA< DENV-1 F 2; DNA; Orthoflavi virus denguei>AC GT T GGAT G GAT T T AGCAACAT T CT RGAT GT CAT GT T< DENV-1 R 1; DNA; Orthoflavi virus denguei>AC GT T GGAT G GAAAC C CAAT ACAT T T CAT GAGT AGAAT T< DENV-1 R 2; DNA; Orthoflavi virus denguei>AC GT T GGAT G T ACAT T T CAT GRGT RGAAT T T CT T GAG< DENV-l-2-3 F; DNA; Orthoflavi virus denguei>ACGTTGGATG AGATYTCTGATGAAYAACCAACG< DENV-2; DNA; Orthoflavi virus denguei>TGGTAGACAGAGGATGGGG< DENV-2 Fl; DNA; Orthoflavi virus denguei>ACGTTGGATG TGCCCAACACAAGGRGAACC< DENV-2 R1; DNA; Orthoflavi virus denguei>ACGTTGGATG GCRCAGGTCACAATGCCYCC< DENV-2 R; DNA; Orthoflavi virus denguei>ACGTTGGATG TGCAGCATTCCAAGTGAGAATCT< DENV-3; DNA; Orthoflavi virus denguei>AAC GC GT GAGAAAC C GT GT GT< DENV-3 Fl; DNA; Orthoflavi virus denguei>ACGTTGGATG CGGGAAAACCGTCTATCAATATGC< DENV-3 R1; DNA; Orthoflavi virus denguei>ACGTTGGATG TGAGAATCTCTTCGCCAACTGTG< DENV-4; DNA; Orthoflavi virus denguei>GGGAAAGGACCCTTACGGATGGT< DENV-4 Fl; DNA; Orthoflavi virus denguei>ACGTTGGATG GGTGAAGAGATTCTCRACYGGACT< DENV-4 R1; DNA; Orthoflavi virus denguei>ACGTTGGATG TGCTGTTGGYGGGATGGAAA< DENV-4 F; DNA; Orthoflavi virus denguei>ACGTTGGATG GGAAGCTTGCTTAACACAGTTCT< DENV-4 R; DNA; Orthoflavi virus denguei>ACGTTGGATG GAGAATCTCTTCACCAACCCTTG< EEEV ext 1; DNA; Alphavirus madariaga>TGCACCCGGACCATCCGA< EEEV F 1; DNA; Alphavirus madariaga>ACGTTGGATG ACACCGCACCCTGATTTTACA< EEEV R 1; DNA; Alphavirus madariaga>ACGTTGGATG CTTCCAAGTGACCTGGTCGTC< Ehrlichia canis 16S ext 1; DNA; Ehrlichia canis> AtCCTCTGGCTATAGG< Ehrlichia chaffeensis 16S ext 1; DNA; Ehrlichia chaffeensis> GTCGAACGGACAATTGCTTATAAC< Ehrlichia chaffeensis 16S F 1; DNA; Ehrlichia chaffeensis> ACGTTGGATG GCGGCAAGCCTAACACATG< Ehrlichia chaffeensis 16S R 1; DNA; Ehrlichia chaffeensis> ACGTTGGATG CCCGTCTGCCACTAACAATTATT< Ehrlichia chaffeensis amp 1; DNA; Ehrlichia chaffeensis>AAT AAAAGAT T TGCAAGATGTTGCGAGTCATGAATCTGGT GT AT CAGATCAGCCAGCTCAAGT T GT TACA G AAAGAGAAAAT GAAAT T GAAT C C CAT CAAGGAGAAACAGAAAAAGAAAGT GGAATAAC T GAAT C T CAT C AGAAAGAAGATGAAATAGTATCTCAACCTTCATCAGAGCCATTTGTTGCAGAAAGTGAAGTTTCTAAAGT T GAAC AAGAAGAAAC T AAC C C T GAAGT T T T AAT AAAAGAT T T G CAAGATGTTGCGAGTCATGAATCTGGTGT AT CAGATCAGCCAGCTCAAGT T GT T ACAGAGAGAGAAAGT GAAAT T GAAT C C CAT CAAGGAGAAACAG AAAAAGAAAGT GGAATAACT GAAT CT CAT CAGAAAGAAGAT GAAAT AGTAT CT CAACCTT CAT CAGAGCC AT T T GT T GCAGAAAGT GAAGT T T CT AAAGT T GAACAAGAAGAAACT AAC C CT GAAGT T T T AAT AAAAGAT T TGCAAGATGTTGCGAGTCATGAATCTGGT GTAT CAGATCAGCCAGCTCAAGTT GTTACAGAGAGAGAAA GT GAAAT T GAAT C C CAT CAAGGAGAAACAGAAAAAGAAAGT GGAATAACT GAAT CT CAT CAGAAAGAAGA TGAAATAGTATCTCAACCTTCATCAGAGCCATTTGTTGCAGAAAGTGAAGTTTCTGAAGTTGAACAAGAA< Ehrlichia chaffeensis amp 2; DNA; Ehrlichia chaffeensis>AT C TAG AAC AAT C T TCTAATTCTGATTTACACGGGTCTTT TTCTGTTGAGTTATTT GAT C C T T T T AAAGA AGCAGTTCAATTGGGGAATGATCTACAACAATCTTCTGATTCTGATTTACACGGGCCTTTTTCTGTTGAG TTATTTGATCCTTCTAAAGAAGAAGTTCAATTGGAGAGTGATCTACCACCATCTTCTAATTCTGATTTAC ACGAGTCTTCTTTTGTTGAGTTACCTGGTCCTTCCAAAGAAGAAGTTCAATTCGAAGATGATGCTAAAAA< Ehrlichia chaffeensis amp 3; DNA; Ehrlichia chaffeensis> AAACTTGAGAGTTTGATCCTGGCTCAGAACGAACGCTGGCGGCAAGCCTAACACATGCAAGTCGAACGGA CAATTGCTTATAACCTTTTGGTTATAAATAATTGTTAGTGGCAGACGGGTGAGTAATGCGTAGGAATCTA CCTAGTAGTATGGAATAGCCATTAGAAATGATGGGTAATACTGTATAATCCCTGCGGGGGAAAGATTTAT CGCTATTAGATGAGCCTACGTTAGATTAGCTAGTTGGTAAGGTAATGGCTTACCAAGGCTATGATCTATA< Ehrlichia chaffeensis TRP120 ext 1; DNA; Ehrlichia chaffeensis> ATCAGCCAGCTCAA ~ ~< Ehrlichia chaffeensis TRP120 F 1; DNA; Ehrlichia chaffeensis>AC GT T GGAT G AAGAT GT T GC GAGT CAT GAAT CT G< Ehrlichia chaffeensis TRP120 R 1; DNA; Ehrlichia chaffeensis> ACGTTGGATG TCCACTTTCTTTTTCTGTTTCTCCTT< Ehrlichia chaffeensis vlpt ext 1; DNA; Ehrlichia chaffeensis> TTGAGTTACCTGGTC< Ehrlichia chaffeensis vlpt F 1; DNA; Ehrlichia chaffeensis> ACGTTGGATG CTAATTCTGATTTACACGAGTCTTC< Ehrlichia chaffeensis vlpt R 1; DNA; Ehrlichia chaffeensis> ACGTTGGATG GCATCATCTTCGAATTGAACTTC< Ehrlichia ewingii amp 1; DNA; Ehrlichia ewingii> CTTTTCTGCAAAAGAAGAAAAAGCAGAGACTAAAAAAACATTTGGTTTAGAAAAAAATTATGATGGAGCT AAAAT AGAAGAT AAT GAAGT ACAGAACAAAT T T AC CAT T T GAAAT TACT CAT T T AAAT AT GAAGAC AAC C CATTTTTAGGTTTTGCTGGAGCCATTGGATATTCAATGGAAGGTCCAAGAATAGAACTTGAAGTATCTTA C GAAACAT T T AAT GT AAAAAAC CAAGACAACAGT T ACAAAAAT GAT GC C CATATGTATTACCTTTTGGCA CGAGAAGTTGATAGTTCTTCGCCAACAAAACCTCAAGTTAACAAATCTGTCTTGCTCAAAAATGAAGGTC TAACTGACTTTTCAATCATGCTAAATGCATGTTATGACATAATAACAGATAATATACCTTTTTCCCCTTA< Ehrlichia ewingii EEM ext 1; DNA; Ehrlichia ewingii>TGG ATA TTC AAT GGA AGG< Ehrlichia ewingii EEM F 1; DNA; Ehrlichia ewingii>ACGTTGGATG G GAG CT AAAAT AGAA GATAATC< Ehrlichia ewingii EEM R 1; DNA; Ehrlichia ewingii>ACGTTGGATG GTGCCAAAAGGT” AATACAT< Ehrlichia muris ext 1; DNA; Ehrlichia muris>ACCCAGCTAATAGTA- < Ehrlichia muris eauclairensis amp 1; DNA; Ehrlichia muris>AT GTTAT AT AAAAT GAGGATAAATAT GTTACAGATAT CT GAAAGGAAT CT GCAAGGAT CT CCT CAGATT G AAT T AC C T AAT T C T T C T C AAGAGAT TCAGTTGTCT GAT AT T GAT AAAAGAGT CAGT GT T GAT C C GTAT GA GTTAGGGTTGCTATTGGCTGGGTTTTTTAGTGCAATGAATTCTTTCTCTTATAGTTGTTCGCAGTATCAT CAT T GT TAT TAT TAT GAT T GT T GT TAT GAT C CT AGT T GT TAT T T T GAT T AT ACT GCT AGT GAT T GT T GT G< Ehrlichia muris ext 2; DNA; Ehrlichia muris>ATTCAGCCAATAGCA- < Ehrlichia muris F 1; DNA; Ehrlichia muris>AAA GAG TCA GTG~ TTG ATC CG< Ehrlichia muris F 2; DNA; Ehrlichia muris>ACGTTGGATG ATG ACAA ATTAG CTGA ATTCAATATTGATCCG< Ehrlichia muris R 1; DNA; Ehrlichia muris>CAA TGA TGA TAC~ TGC GAA CA< Ehrlichia muris R 2; DNA; Ehrlichia muris>ACGTTGGATG TGA TAA TGA TAG TGC GGA CA< Ehrlichia ruminantium F 1; DNA; Ehrlichia ruminantium>ACGTTGGATG CAA AAC TAG TAG AAA TTG CAC A< Ehrlichia ruminantium R 1; DNA; Ehrlichia ruminantium>ACGTTGGATG TGC ATC TTG TGG TGG TAG< Ehrlichia spp 16S ext 1; DNA; Ehrlichia sp. >CCT GTT TGC TCC CCA CGC T< Ehrlichia spp 16S ext 2; DNA; Ehrlichia sp. >CTTACTAGTCCACCTACGTG< Ehrlichia spp 16S F 1; DNA; Ehrlichia sp. >ACGTTGGATG TAG AAA GAT AAA ATC CTC ACA TTT< Ehrlichia spp 16S F 2; DNA; Ehrlichia sp. >ACGTTGGATG TCG ACC TCT AGT CTA GCA GTA TTA< Ehrlichia spp 16S R 1; DNA; Ehrlichia sp. >ACGTTGGATG ATT CGT AGA TAT TAG GAG GAA CAC< Ehrlichia spp 16S R 2; DNA; Ehrlichia sp. >ACGTTGGATG CGG AAT TAT TGG GCG TAA AG< Ehrlichia spp amp 1; DNA; Ehrlichia sp. > TGACGGTACCTATAGAAGAAGTCCCGGCAAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGGGCAAGC GTTGTTCGGAATTATTGGGCGTAAAGGGCACGTAGGTGGACTAGTAAGTTAAAAGTGAAATACCAAAGCT CAACTTTGGAGCTGCTTTTAATACTGCTAGACTAGAGGTCGAAAGAGGATAGCGGAATTCCTAGTGTAGA< Ehrlichia spp amp 2; DNA; Ehrlichia sp. > GGTGAAATTCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTATCTGGTTCGATACTGACACTGA GGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGAGTGCTAA ATGTGAGGATTTTATCTTTGTATTGTAGCTAACGCGTTAAGCACTCCGCCTGGGGACTACGGTCGCAAGA< Ehrlichia theileri ext 1; DNA; Ehrlichia sp. > AGTACAACCAGCTCCTGCT< Francisella tularensis 23S ext 1; DNA; Francisella tularensis> CAGTTCTCACATGAATGGTCTCGC< Francisella tularensis 23S F 1; DNA; Francisella tularensis> ACGTT GGAT G T GAGAT GATAACAAGACAACAGGTAACA< Francisella tularensis 23S R 1; DNA; Francisella tularensis>AC GT T G GAT G G GAT GAGAT C CT AT AC AT G C AGT AG G< Francisella tularensis amp 1; DNA; Francisella tularensis> AGAGCACAT GCTT GT GCTACAGGATAT GATAAAGAT GAT AAT CAAGCAATTGGTAGATCAGTTGGTGGGA TAACCACTAAAATCCATGCTATGACTGATGCTTTAGGTAATCCAATAGAAATATTGTTGTCAGAAGGTAA< Francisella tularensis amp 2; DNA; Francisella tularensis>T T GAAC C AGAAT T AT T C G GT AC AAAT AAT AAC T AAAAAAAG GAGAAT GAT TAT GAGT GAGAT GAT AACAA GACAACAGGTAACAAGT GGCGAGACCATTCATGTGAGAACTGAT CCTACT GCAT GTATAGGAT CT CAT CC TAATTGTAGATTGTTTATTGATTCTTTAACTATAGCTGGGGAGAAACTTGATAAAAATATCGTTGCTATA< Francisella tularensis amp 3; DNA; Francisella tularensis> AGTTATTACCTTGCTTAACTGTTACAGTTGCCCAAGTTTTATCGTTCTTCTCAGCATACTTAGTAATTGG GAAGCTTGTATCATGGCACTTAGAACCTTCTGGAGCCTGCCATTGTAATCTTACACTTCCTTGTGGGTTA< Francisella tularensis ISFtu2 ext 1; DNA; Francisella tularensis> CCATGCTATGACTGATGCTTTAGG< Francisella tularensis ISFtu2 F 1; DNA; Francisella tularensis> ACGTTGGATG TTGGTAGATCAGTTGGTGGGATAAC< Francisella tularensis ISFtu2 R 1; DNA; Francisella tularensis> ACGTTGGATG TGAGTTTTACCTTCTGACAACAATATTTC< Francisella tularensis tul4 ext 1; DNA; Francisella tularensis> GTGCCATGATACAAGCTTCCC “ “ “< Francisella tularensis tul4 F 1; DNA; Francisella tularensis> ACGTTGGATG ATTACAATGGCAGGCTCCAGA< Francisella tularensis tul4 R 1; DNA; Francisella tularensis> ACGTTGGATG TGCCCAAGTTTTATCGTTCTTCT< Heartland virus ext 1; DNA; Heartland bandavirus> AGTGGGAAACCGTCAGT< Heartland virus F 1; DNA; Heartland bandavirus>ACGTTGGATG AGCAGGAGAAGAGTGCATTG< Heartland virus R 1; DNA; Heartland bandavirus>ACGTTGGATG ACTGCCCAGGTGTTCTTG< Heartland virus S ext 1; DNA; Heartland bandavirus> TGGATGCCTATTCCCTTTGG< Heartland virus S F 1; DNA; Heartland bandavirus>ACGTTGGATG CCTTTGGTCCACATTGATTG< Heartland virus S R I; DNA; Heartland bandavirus>ACGTTGGATG CACTGATTCCACAGGCAGAT< JEV ext 1; DNA; Orthoflavi virus j aponicum>CAAGAGGTGGACGGC< JEV F 1; DNA; Orthoflavi virus j aponicum>ACGTTGGATG GCCACCCAGGAGGTCCTT< JEV R 1; DNA; Orthoflavi virus j aponicum>ACGTTGGATG CCCCAAAACCGCAGGAAT< LACV ext 1; DNA; Orthobunyavirus lacrosseense> TGTGCAAGTCGAAAGGGCCTGC< LACV F 1; DNA; Orthobunyavirus lacrosseense>ACGTTGGATG TATAAAAGCCTAAGAGCTGCCAGAGT< LACV R 1; DNA; Orthobunyavirus lacrosseense>ACGTT GGAT G GAC CAGT ACT GCAGT AAT T AT AGACAAT< P falciparum ext 1; DNA; Plasmodium falciparum>CT AAAAGT CAC CT C GAAAGAT GAC< P falciparum F 1; DNA; Plasmodium falciparum>ACGTTGGATG CCGACTAGGTGTTGGATGAAAGTGTTAA< P falciparum R 1; DNA; Plasmodium falciparum>ACGTTGGATG AACCCAAAGACTTTGATTTCTCATAA< P knowlesi ext 1; DNA; Plasmodium knowlesi>catCGGAGGCATCAGTTA< P knowlesi F 1; DNA; Plasmodium knowlesi>ACGTTGGATG CTAAAATGCGCACAAAGTCGAT< P knowlesi R 1; DNA; Plasmodium knowlesi>ACGTTGGATG GCAGTTAAAACGCTCGTAGTTGAA< P malariae ext 1; DNA; Plasmodium malariae>gccCTATCTAAAA GAAACACTCA< P malariae F 1; DNA; Plasmodium malariae>ACGTT GGAT G~ CCGACTAGGT GTT GGAT GAT AGAGTAAA< P malariae F 2; DNA; Plasmodium malariae>ACGTT GGAT G~ CGACTA GGT GTT GGAT GATAGAGTA< P malariae R 1; DNA; Plasmodium malariae>ACGTTGGATG AACCCAAAGACTTTGATTTCTCATAA< P ovale ext 1; DNA; Plasmodium ovale>cggCGAAAGGAATTTTC< P ovale F 1; DNA; Plasmodium ovale>ACGTTGGATG CCGACTAGGTTTTGGATGAAAGATTTTT< P ovale R 1; DNA; Plasmodium ovale>ACGTTGGATG AACCCAAAGACTTTGATTTCTCATAA< P vivax ext 1; DNA; Plasmodium vivax>CT AAGAAT AAACT C C GAAGAG< P vivax F 1; DNA; Plasmodium vivax>ACGTTGGATG CCGACTAGGCTTTGGATGAAAGATTTTA< P vivax R 1; DNA; Plasmodium vivax>ACGTTGGATG AACCCAAAGACTTTGATTTCTCATAA< Panola Mountain Ehrlichia ( PME) amp 1; DNA; Ehrlichia> TAATGTCATTTCCACAGCATTCTCATCCTATGGCTATGTTAATAGCGTGTTTTTCTGCCTTAGCTGCACA TTATTGTGATAAGGAAACTAGTTATGAGTTAGAATGTAAACTTGCTATCGCTAAAATTGCAAGTATGATT GCGCTAATTTATAGGTATACTACTAATCAAGATTTTATTCAAGCTGATTCAAGATTATCCTATAGTAAAA< Panola Mountain Ehrlichia ( PME) amp 2; DNA; Ehrlichia sp. > TACATGTTCTATTTTAACTGCTAAAGTTATTGAGGAAGTATCTAAGGCTAAAGCTGCTGGTGCAGATATT GTTT GTATTAAAGAAGGT GTACTTAAAGCTAAGGAGGCAGTATTAGAAGCTTTAAT GT CTAT GAAACGT G AAATATTGTCTGAAGAAGAAATTGCTCAAGTTGCAACTATTTCTGCTAATGGAGATAAGAATATAGGGGC< Panola Mountain Ehrlichia gltA ext 1; DNA; Ehrlichia sp. > TGCCTTAGCTGCACATTATTGTGA< Panola Mountain Ehrlichia gltA F 1; DNA; Ehrlichia sp. >AC GT T GGAT G T GT CAT T T C CACAGCAT T CT CAT C< Panola Mountain Ehrlichia gltA R 1; DNA; Ehrlichia sp. > ACGTTGGATG ATTAGCGCAATCATACTTGCAA< Panola Mountain Ehrlichia groEL290 ext 1; DNA; Ehrlichia sp. > GCTGCTGGTGCAGATATTGTT ~ ~< Panola Mountain Ehrlichia groEL290 F 1; DNA; Ehrlichia sp. > ACGTTGGATG GTTATTGAGGAAGTATCTAAGG< Panola Mountain Ehrlichia groEL290 R 1; DNA; Ehrlichia sp. > ACGTTGGATG TTA AAG CTT CTA ATA CTG~CCT C< Plasmodium sp ext 1; DNA; Plasmodium sp. >AAACGGCCATGCATCACCA< Plasmodium Sp F 1; DNA; Plasmodium sp. >ACGTTGGATG” AGCTCTTTCTTGATTTCTTGG< Plasmodium sp R 1; DNA; Plasmodium sp. >AC GT T GGAT G~GT T AAGGGAGT GAAGAC GA T CAGA< Plasmodium Sp R 1; DNA; Plasmodium sp. >ACGTTGGATG CAGACAAATCATATTCACGAACT< Plasmodium sp R 2; DNA; Plasmodium sp. >AC GT T G GAT G~AAC C C AAAGAC T T T GAT T T C T C AT AA< Powassan virus NS5 ext 1; DNA; Powassan virus> AGGGTGAGGATGTGCACCA< Powassan virus NS5 ext 2; DNA; Powassan virus>CCAAAGTGAGGATGTGTAC< Powassan virus NS5 F 1; DNA; Powassan virus>AC GT T GGAT G GAT CAT GAGAGC G / TGT GAGT GACT< Powassan virus NS5 R 1; DNA; Powassan virus>ACGTTGGATG GGAT / CCTCACCTT T / CGCTATGAAT / CTCA< Powassan virus UTR ext 1; DNA; Powassan virus> TACTGCGGCAGCACACACAGT< Powassan virus UTR ext 2; DNA; Powassan virus> aACTGCGGCAGCACACTCAGT< Powassan virus UTR F 1; DNA; Powassan virus>ACGTTGGATG G / ATGA / GTGTGGCAGCGCAC< Powassan virus UTR R 1; DNA; Powassan virus>ACGTTGGATG CTGCGTCGGGAGCGACCA< Rickettsia 364D ext 1; DNA; Rickettsia sp. >ATG CGT CGT CAT~ AGC~ CGT A< Rickettsia 364D amp 1; DNA; Rickettsia sp. > TAAAAGTTGGACTAAATACGGTTTATTAATTACTTAAGCTGTATTGTACGAATGTTCAGGATGTCATTGT TGCGGGATAATGCGTCGTCATAGCCGTAGGCTGCGTGGCAATCTAGGAGAAATAATAAAAAACTTCTGAT TTACAGAATTTTTTTCTGCCTTGTTTTGTCAAAACTTACAGCTTTTCCTTGCAATGATGGGAAACAAGAT< Rickettsia aeschlimannii amp 1; DNA; Rickettsia aeschlimannii>T GATCTTGCTGCTGCCAATAATTGGAATGATATAACGGCTGAAGGGATAGCTAATGGTACTCCTGTTGAT GGTCCTCAAAATGGTATGGCATTTACTTACGGTGGT GAT CATACTAT CACT GCAGAT GAAGCCGGT CGTA< Rickettsia 364D F 1; DNA; Rickettsia sp. >ACGTTGGATG AGG ATG TCA TTG TTG CGG GA< Rickettsia 364D R 1; DNA; Rickettsia sp. >ACGTTGGATG TCT CCT AGA TTG CCA CGC AG< Rickettsia aeschlimannii amp 1; DNA; Rickettsia aeschlimannii> TGATCTTGCTGCTGCCAATAATTGGAATGATATAACGGCTGAAGGGATAGCTAATGGTACTCCTGTTGAT GGTCCTCAAAATGGTATGGCATTTACTTACGGTGGTGATCATACTATCACTGCAGATGAAGCCGGTCGTA< Rickettsia aeschlimannii ext 1; DNA; Rickettsia aeschlimannii> CTAATGGTACTCCTGTTGAT GG< Rickettsia aeschlimannii F 1; DNA; Rickettsia aeschlimannii>AAT GATATAACGGCT GAAG< Rickettsia aeschlimannii R 1; DNA; Rickettsia aeschlimannii> TGGTATGGCATTTACTTAC< Rickettsia africae ext 1; DNA; Rickettsia africae> CTAATGGTACTCCTGTTGATGG- < Rickettsia africae F 1; DNA; Rickettsia africae>ACGTTGGATG AAT GATATAACGGCT GAAG< Rickettsia africae R 1; DNA; Rickettsia africae>ACGTTGGATG GTAAGTAAATGCCATACCA;< Rickettsia afzelii tsaB R 1; DNA; Rickettsia sp. >ACGTTGGATG ATTCTTGTGGTCCTGGTT< Rickettsia afzelii tsaB F 1; DNA; Rickettsia sp. >ACGTTGGATG TGAATCAATCTGCCCTAG< Rickettsia akari amp 1; DNA; Rickettsia akari>GGTTCTGTCTTTAAACTTGCTGACGATACAGTTATAAACGGTAAAGTTAACGAAAATGCTCTTATTGGTG GTGCTGTTGCAGGTGGTACTATTCAGTTAGATGGAAGTGCTACAATTACCGGTGATATAGGTAACGGTGC TGGTAATGCTGCATTACACGGTATTACTTTAGCTGATGATGCTTCAAAAACATTAACTCTCGGTGGAGCA AAGATTATCGGTGTCAATGCCGGTGGAACGATTAATTTCCAAGCTAACGGTGGTACTATTAAATTAACAA< Rickettsia akari ompB ext 1; DNA; Rickettsia akari> GTGCTGGTAATGCTGCA< Rickettsia akari ompB F 1; DNA; Rickettsia akari>ACGTTGGATG GTG GTG CTG TTG GAG GTG G< Rickettsia akari ompB R 1; DNA; Rickettsia akari>ACGTTGGATG TTG CTC CAC CGA GAG TTA ATG TT< Rickettsia amblyommatis amp 1; DNA; Rickettsia sp. > CCTTGACCGTCACCGATATTAATTGCCTTAACAGTAGCAAAACTTTTATCTGAAACTTGAATAAATCCAT TAGTAACATTTAATGTACCGTTAACACCATTAACTATTAAAGCATTCTGTCCAAGTGTAAGAGGAGCCTT TTGAGTTGTAGGATTTGCTAAATTAAATACTAATGTAGAAGCCGCAGCACCAAAGTCTATTGTTCCTACA< Rickettsia amblyommatis ompB ext 1; DNA; Rickettsia sp. > CGCGATCTCCTCTTACACTTG - - - < Rickettsia amblyommatis ompB F 1; DNA; Rickettsia sp. >ACGTTGGATG GGT GCT CGC GCT TCT~ ACA TTA G< Rickettsia amblyommatis ompB R 1; DNA; Rickettsia sp. >ACGTTGGATG CTG AAA CTT GAA TAA"~ATC CAT TAG TAA CAT< Rickettsia bellii amp 1; DNA; Rickettsia bellii> GCTATTATGCTTGCGGCGGTTGGTTCTCTTTCTGCATTTTATCCTGATTTGCTGAATTTTTTTAAAGAAG CGGATTATGAACTTACAGCTATTAGAATGATAGCTAAAATACCAACTATTGCTGCAATGTCTTATAAATA TTCTATAGGTCAGCCTTTTGTTTATCCCGATAATTCTTTAGATTTTACTGAAAATTTCTTGCATATGATG TTTGCCACACCTTGTGAAAAATATAAAGTAAATCCAGTAATAAAAAATGCCTTAAACAAGATATTTATTT TACATGCTGATCATGAGCAAAATGCTTCTACTTCAACAGTCAGAATTGCCGGCTCTTCAGGAGCTAACCC TTTTGCTTGCGTCAGTACTGGTATTGCATCGCTTTGGGGTCCTGCTCATGGCGGGGCTAATGAAGCAGTG< Rickettsia bellii git ext 1; DNA; Rickettsia bellii>G ATG TTT GCC ACA CCT TGT GA< Rickettsia bellii git F 1; DNA; Rickettsia bellii>ACGTTGGATG ATC CTG ATT TGC TGA ATT TTT T< Rickettsia bellii git R 1; DNA; Rickettsia bellii>ACGTTGGATG TGC AAT ACC AGT ACT GAC G< Rickettsia CA6267 ext 1; DNA; Rickettsia sp. >CTAATGGTGATCCTACTGGC< Rickettsia CA6267 F 1; DNA; Rickettsia sp. >ACGTTGGATG GGGCACTTGGTGTTCCTACA-3< Rickettsia CA6267 R 1; DNA; Rickettsia sp. >ACGTTGGATG AAATGCCCAATTGTTTTGAGGAC-3< Rickettsia canis 16S F 1; DNA; Rickettsia sp. >ACGTTGGATG AGCYTAACACATGCAAGTCGAA< Rickettsia canis 16S R 1; DNA; Rickettsia sp. >ACGTTGGATG TTACTCACCCGTCTGCCACTA7\< Rickettsia conorii amp 1; DNA; Rickettsia conorii> CAAGGTCTTAAAGCCGCTTTATTCACCACCTCAACCGCAGCGATAATGCTGAGTAGTAGCGGGGCACTCG GTATTGCTGTTTCAGGTGTTATTGCTACTA ATAATAATGCAGCATTTAGTGATAATGTTGGCAATAATTG GAATGAGATAACGGCTGCAGGGGTAGCTAATGGTACTCCTGCTCGCGGTCCTCAAAACAATTGGGCATTT ACTTACGGTGGTGATTATACTATCACTGCAGATGTAGCCGATCATATTATTACGGCTATAAATGTTGCGG< Rickettsia conorii ompA ext 1; DNA; Rickettsia conorii> TGGAATGAGATAACGGCTG< Rickettsia conorii ompA F 1; DNA; Rickettsia conorii>ACGTTGGATG CGGGGCACTCGGTATTGCTGTTT< Rickettsia conorii ompA R 1; DNA; Rickettsia conorii>ACGTTGGATG GCGAGCAGGAGTACCATTAGC< Rickettsia Helvetica 23S ext 1; DNA; Rickettsia helvetica>AAC CGT AGC GTA CAC “ “ “< Rickettsia Helvetica 23S F 1; DNA; Rickettsia helvetica> ACGTTGGATG TTT GAA GGA GAC~ACG GAA CAC A< Rickettsia Helvetica 23S R 1; DNA; Rickettsia helvetica> ACGTTGGATG TCC GGT ACT CAA"~ATC CTC ACG TA< Rickettsia montanensis amp 1; DNA; Rickettsia montanensis>AGGTCTGGGTCTTGGAAGCGATAACGGCATAATCGTTAATGCTACTAAATTATATGCAGGTATCGGTACTACAAACAATAATCAAGGTACTGTCATACTTAGCGGTGGTGTTCCTAATACCCCTGGTACAGTTTATGGCT TAGGCAAAGATGCTAGCGCTTCACAGTTAAAGCAAGTAACGTTCACTACAGACTATAACAACTTAGGTAA TATTATTGCAACTAACACAACAATTAATGACGGTGTAACTGTTACTACAGGCGGTATAGCCGGAACAGTT TTTAACGGTAAAATTACTCTTGGAAGTGTTAACGGTAACGCTAATGTAAGATTTGCTGACGGTATATTGT< Rickettsia montanensis ompB ext 1; DNA; Rickettsia montanensis> AT GCT AGC GCT TCA CA ~< Rickettsia montanensis ompB F 1; DNA; Rickettsia montanensis> ACGTTGGATG GCG GTG GTG~ TTC CTA ATA C< Rickettsia montanensis ompB R 1; DNA; Rickettsia montanensis> ACGTTGGATG CCT AAG TTG~ TTA TAG TCT GTA GTG< Rickettsia parkeri amp 1; DNA; Rickettsia parkeri> GATGGTGCGGGATTTGACCAAACTGCAGTTCCTGCAAATGTTGCAGTTCCTCTAAATGCAGTTATTACTG CTGGTGTTCATAGGGGTATTAATTTAAATACTCCAGCCGGTAGTTTTAACGGTTTGTTTTTAAATGCTGC AAACAATTTAGCAGTGACAGTGAGTGAAGATACTACCTTAGGGTTCATCACTCGTGCTGTTAATAACGCT< Rickettsia parkeri ompB ext 1; DNA; Rickettsia parkeri> TACCCTTAGAGCAGCAGC”< Rickettsia parkeri ompB ext 2; DNA; Rickettsia parkeri> TACCCCTATGAACACCAGC< Rickettsia parkeri ompB F 1; DNA; Rickettsia parkeri>ACGTTGGATG CAA ATG~ TTG CAG TTC CTC TAA ATG< Rickettsia parkeri ompB R 1; DNA; Rickettsia parkeri>ACGTTGGATG AAA ACA"~AAC CGT TAA AAC TAG CG< Rickettsia prowazekii amp 1; DNA; Rickettsia prowazekii> CACTACTTGGTAGCTCCCCATAAATCATCAAATATGCCACTTCTAAAAAATCACTTTTCTCAGCTAAGTC T T TAATATCATATCCTCGATACCATAATATGCCTT TAT CAC C GT CT AT AT AT GT GAT AGT AGAT T GACAA GAAGCAGTAGACATAAAACCCGGATCATAAGTAAAGTAATCGGCTTCCGCAGATACCCTACTTATATCGA TTACATCTTTACCGATACTTGCTTTAAGTATAGGTAACTTAAATAGTTTACCTCTAATTTTTAATTCTGC< Rickettsia prowazekii gltA ext 1; DNA; Rickettsia prowazekii> cgTACTTATGATCCGGGT< Rickettsia prowazekii gltA F 1; DNA; Rickettsia prowazekii> ACGTTGGATG TCGGTAAAGATGTAATCGATATAAG< Rickettsia prowazekii gltA R 1; DNA; Rickettsia prowazekii>CAT AT C CTC GAT AC CAT AAT AT GC< Rickettsia rickettsia ext 1; DNA; Rickettsia rickettsii>CC TCT CCA ATC AGC GAT TC~< Rickettsia rickettsia 0 F 1; DNA; Rickettsia rickettsii> ACGTTGGATG AAA TCA ACG~ GAA GAG CAA AAC< Rickettsia rickettsia 0 F 2; DNA; Rickettsia rickettsii> ACGTTGGATG AAA TCA ACG GAA GAG CAA AAC< Rickettsia rickettsia 0 R 1; DNA; Rickettsia rickettsii> ACGTTGGATG CCC TCC ACT~ACC TGC ATC AT< Rickettsia rickettsia 0 R 2; DNA; Rickettsia rickettsii> ACGTTGGATG CCC TCC ACT ACC TGC ATC AT< Rickettsia rickettsia 23S ext 1; DNA; Rickettsia rickettsii> CGGATCGAAGTTTATTCGCA “ “ “< Rickettsia rickettsia 23S F 1; DNA; Rickettsia rickettsii> ACGTTGGATG GCGATGAAGGACGTAATACGCT< Rickettsia rickettsia 23S R 1; DNA; Rickettsia rickettsii> ACGTTGGATG TAGGTAGGTTTCCCTATTCGGA< Rickettsia rickettsia amp 1; DNA; Rickettsia rickettsii>GACAAAGT T T GGAAGAGAGTAAAT CAACGGAAGAGCAAAACT T T CAAT CAGAAGGAGT GGT AAGT T CAGT AAGTAAGAT GCCAACCATAAATT CTAGT GAGTCCTCTCCAATCAGCGATT CAGGCAATAAGT CAGACT CT GACT CAAAAGAT GATGATGCAGGTAGTGGAGGGGTTAAT CAAT CTT GACAAAAATT GGCT GGT GT GATTA GCAAAATTGCCAGCAAGGTTAAAGGTTGGGGTTTGCCTCAGAAGTTAGAGGAGTTTCACCCATTGACTGA< Rickettsia rickettsia amp 2; DNA; Rickettsia rickettsii>GACAAAGT T T GGAAGAGAGTAAAT CAACGGAAGAGCAAAACT T T CAAT CAGAAGGAGT GGT AAGT T CAGT AAGTAAGATGCCAACCATAAATTCTAGTGAGTCCTCTCCAATCAGCGATTCAGGCAATAAGTCAGACTCT GACT CAAAAGAT GATGATGCAGGTAGTGGAGGGGTTAAT CAAT CTT GACAAAAATT GGCT GGT GT GATTA GCAAAATTGCCAGCAAGGTTAAAGGTTGGGGTTTGCCTCAGAAGTTAGAGGAGTTTCACCCATTGACT< Rickettsia rickettsia ompB ext 1; DNA; Rickettsia rickettsii> GTTCCTAATGCTATAAC - - - < Rickettsia rickettsia ompB F 1; DNA; Rickettsia rickettsii>ACGTTGGATG ATAACCCAAGACTCAAACTTTGGTA< Rickettsia rickettsia ompB R 1; DNA; Rickettsia rickettsii> ACGTTGGATG GCAGTGTTACCGGGATTGCT< Borrelia rickettsii ext 2; DNA; Borrelia sp. >TCC TCT CCA ATC AGC~ G< Rickettsia ruminantium groEL 1 amp 1; DNA; Rickettsia sp. > AGTTGCAACTTGGGCAATTTCTTCTTCAGATAACACTTCACGCTTCATGCATTTTAAAGCTTCTAATACTGCCTC TTTAGCTTTAAGTACACCTTCCCTAACACAAATAATATCTGCACCAGCAGCCTTAACCTTAGATA CTTCCTCAATAACCTT T GCAGT T AAAAT AGAACAT GT AGT AGT T C CAT CAC CT ACT T TAT C GT T ACAT T G< Rickettsia ruminantium groEL 1 ext 1; DNA; Rickettsia sp. > TACACCTTCCCTAACACAAATAATATCTGCAC< Rickettsia ruminantium groEL 1 F 1; DNA; Rickettsia sp. > TTAAAGCTTCTAATACTGCCTC ~ ~< Rickettsia ruminantium groEL 1 R 1; DNA; Rickettsia sp. > CCTTAGATACTTCCTCAATAAC< Rickettsia ruminantium groEL 2 ext 2; DNA; Rickettsia sp. > GTACACCTTCCCTAAC< Rickettsia spp ext 1; DNA; Rickettsia sp. >CCT GCT TCT ATT TGT CTT GCA GT< Rickettsia spp 0 F 1; DNA; Rickettsia sp. >ACGTTGGATG AGC TTG CTT TTG GAT CAT TTG G< Rickettsia spp 0 R 1; DNA; Rickettsia sp. >ACGTTGGATG TTC CTT GCC TTT TCA TAG ATC TAG T< Rickettsia spp amp 1; DNA; Rickettsia sp. > GAACCTTTACCTTTACCCATTCTTACTTCGGCAGGCTTTTGAGAAACGGGAACATCCGGAAAAATGCGAA TCCATAATCTTCCTTGCCTTTTCATACATCTAGTAGCAGCTTTTCTTCCTGCTTCTATTTGTCTTGCAGT AACACGCCAACCATCTATAGATTTTAGACCAAATGATCCAAAAGCAAGCGTCGTACCTGCTTTTGCTGTT GAAGCAACCCTACCCTTATGAGCTTTTCTAAATTTTTGTTTTTTCGGAGCTAACATTTTAATACTTAAAA< Rickettsia spp amp 2; DNA; Rickettsia sp. > TTTTTTGTTCAGGGTCTTCGTGCATTTCTTTCCATTGTGCCATCCAGCCTACGGTTCTTGCTATTGCAAA AAGTACCGTGAACATTTGCGAC G GT AT AC C CAT AG CTT T AT AGAT AAT AC C C GAAT AAAAAT C AAC AT T T GGATATAATTTTCTCTCAATAAAATATTCATCTTTAAGAGCGATAGCTTCAAGTTCTATTGCTATTTGTA< Rickettsia spp git F 1; DNA; Rickettsia sp. >ACGTTGGATG TCG CAA ATG TTC ACG GTA CTT T< Rickettsia spp git R 1; DNA; Rickettsia sp. >ACGTTGGATG TCG~ TGC~ ATT TCT TTC CAT TGT G< Rickettsia spp gltA ext 1; DNA; Rickettsia sp. >GC AAG AAC CGT~ AGG CTG GA< Rickettsia typhi 23S ext 1; DNA; Rickettsia typhi>GCACATTTGACTTCTA- < Rickettsia typhi 23S F 1; DNA; Rickettsia typhi>ACGTTGGATG CTAACGCCTCTGCTTCGCAG< Rickettsia typhi 23S R 1; DNA; Rickettsia typhi>ACGTTGGATG GAAAGACCCCGTGAACCTTTACTA< Rickettsia typhi amp 1; DNA; Rickettsia typhi> GTAACTAATGTTACTAAACAGGGTAACTTCTTTAATTTTACTATTGGTGCTGGTAAAAGTCTTACCATAA CAGGT CATGGTATTACTGCTCAACAAGCTGCTACTACAAAAAGTGCTCAAAAT GTT GTTT CAAAAGTTAA TGCTGGTGCTGCTATTAACGATAATGATCTTAGCGGTGTAGGATCAATAGACTTTACTGCTGCGCCTTCT GTATTAGAATTTAATTTAATAAATCCTACAACTCAAGAAGCTCCTCTTACACTTGGTGATAATGCTAAAA< Rickettsia typhi ompB ext 1; DNA; Rickettsia typhi> CGCGATCGTTAATAGCAGCACCA< Rickettsia typhi ompB F 1; DNA; Rickettsia typhi>ACGTTGGATG TGG TAT TAG TGC TCA ACA AGC T< Rickettsia typhi ompB R 1; DNA; Rickettsia typhi>ACGTTGGATG CAG TAA AGT CTA TTG ATC CTA CAC C< SLEV ext 1; DNA; Orthoflavi virus louisense>GGT GTT CAAAGAAAAGGTT< SLEV NS5 F 1; DNA; Orthoflavi virus louisense>ACGTTGGATG GGTGGTTCGGGAGCCCTT< SLEV NS5 F 2; DNA; Orthoflavi virus louisense>ACGTTGGATG- GGTGGTTCGGGAGCCTTT< SLEV NS5 R 2; DNA; Orthoflavi virus louisense>ACGTTGGATG CACGCCTTTTGGCCAACAA< SLEV NS5 R 3; DNA; Orthoflavi virus louisense>ACGTTGGATG- CACGCCTTTTGGTCAACAA< Tick-borne encephalitis virus ext 1; DNA; Tick-borne encephalitis virus> tCCCATCACTCCAgTgTCA< Tick-borne encephalitis virus ext 2; DNA; Tick-borne encephalitis virus> aCCCATCACTCCTgTgTCA< Tick-borne encephalitis virus ext 3; DNA; Tick-borne encephalitis virus> T GAG C C AC CAT C AC C C AGAC AC< Tick-borne encephalitis virus F 1; DNA; Tick-borne encephalitis virus> AC GT T GGAT G T g gAYT T YAgACAg gAAYCAACACA< Tick-borne encephalitis virus F 3; DNA; Tick-borne encephalitis virus> ACGTTGGATG GGG CGG TTC TTG TTC TCC< Tick-borne encephalitis virus R 1; DNA; Tick-borne encephalitis virus> ACGTTGGATG TCCAgAgACTYTgRTCDgTgTggA< Tick-borne encephalitis virus R 3; DNA; Tick-borne encephalitis virus> ACGTTGGATG ACA CAT / G CAC CTC CTT GTC AGA CT< WEEV ext 1; DNA; Alphavirus western>ATACGGCAATACCACCGCGCAC< WEEV F 1; DNA; Alphavirus western>ACGTTGGATG CTGAAAGTCGGCCTGCGTAT< WEEV R 1; DNA; Alphavirus western>ACGTTGGATG CGCCATTGACGAACGTATCC< WNV ext 1; DNA; Orthoflavi virus nilense>GACCAT GGGAGAAGCT< WNV ext 2; DNA; Orthoflavi virus nilense>AACTATGGGTGAAGCCC< WNV F 1; DNA; Orthoflavi virus nilense>ACGTTGGATG TCAGCGATCTC / TTCCACCAA / GAG< WNV F 2; DNA; Orthoflavi virus nilense>ACGTTGGATG TCAGTGACTTA / GTCAACA / GAGAG< WNV R 1; DNA; Orthoflavi virus nilense>ACGTTGGATG CTGGGTCAGCACGTTTGTCAT< WNV R 2; DNA; Orthoflavi virus nilense>ACGTTGGATG CT / GG GGT CAG CAC G / TTC / T TG / CT CGT< WNV 3-F 1; DNA; Orthoflavi virus nilense>ACGTTGGATG TCAGCGATCT YTCCACCARAG< WNV 3-F 2; DNA; Orthoflavi virus nilense>ACGTTGGATG TCAGTGACTTRTCAACRAGAG< WNV 3-R 1; DNA; Orthoflavi virus nilense>ACGTTGGATG CTGGGTCAGCACGTTTGTCAT< WNV 3-R 2; DNA; Orthoflavi virus nilense>ACGTTGGATGCKGGGT CAG CAC KTY TST CGT< WNV ext 1; DNA; Orthoflavi virus nilense>GACCAT GGGAGAAGCT< WNV ext 2; DNA; Orthoflavi virus nilense>AACTATGGGTGAAGCCC< YFV ext 1; DNA; Orthoflavi virus flavi>GCTAGGCAATAAACACATTTGG< YFV F 1; DNA; Orthoflavi virus flavi>ACGTTGGATG AGGTGCATTGGTCTGCAAAT< YFV mdl F 1; DNA; Orthoflavi virus flavi>ACGTTGGATG ATCGTTCKTTGAGCGATTAGCAG< YFV mdl R 1; DNA; Orthoflavi virus flavi>ACGTTGGATG TGTTTTATTTTGYTTGACAAGGAGCG< YFV R 1; DNA; Orthoflavi virus flavi>ACGTTGGATG TCTCTGCTAATCGCTCAACG< ZIKV ext 1; DNA; Orthoflavi virus zikaense>ggCTCAGACCAGCTGA< ZIKV ext 2; DNA; Orthoflavi virus zikaense>attCAC CAT CCG CAA AGTGC< ZIKV ext 3; DNA; Orthoflavi virus zikaense>atgCAC CAT CCG CAA AGTTC< ZIKV F 1; DNA; Orthoflavi virus zikaense>ACGTTGGATG AARTACACATACCARAACAAAGTG GT< ZIKV R 1; DNA; Orthoflavi virus zikaense>ACGTTGGATG TCCRCTCCCYCTYTGGTCTTG< ZIKV SPOV F 1; DNA; Orthoflavi virus zikaense>ACGTTGGATG TGG AGY TAG TAY GCC GCM< ZIKV SPOV R 1; DNA; Orthoflavi virus zikaense>ACGTTGGATG TGG AGY TAG TAY GCC GCMARTHROPOD IDENTIFICATION SEQUENCES (SEQ ID NOS: 377-418) < Aedes albopictus ext 1; DNA; Aedes albopictus> TCAATTCGCGGCAATCGTAAACCT< Aedes albopictus F 1; DNA; Aedes albopictus>ACGTTGGATG CTCAAGCAGTCGCCAGTCAACAAG< Aedes albopictus R 1; DNA; Aedes albopictus>ACGTTGGATG TCCCTCGAACAACGGCAACTG< Aedes aegypti ext 1; DNA; Aedes aegypti>T CGT CAAAT GGAAT GGGCAAT GGT G< Aedes aegypti F 1; DNA; Aedes aegypti>AC GT T GGAT G T GGCT AGT CT GGAC GAT GAAAGT GAG< Aedes aegypti R 1; DNA; Aedes aegypti>ACGTTGGATG GGTAGGTGGAATTTTGGGATGGTAGTC< Amblyomma Americanum ext 1; DNA; Amblyomma americanum> ACGACGTAACGCGGGACGG< Amblyomma Americanum amp 1; DNA; Amblyomma americanum> GTTTTGACCGTGTCGGCAAAGAGGACGGCTCTGTACGAAACGCCGAAGATCCCTCGAAAGACTTTGCGAA AGAGGAGAGTGGGTCGAACCCTTCAGCCGCGCTCCAAGCGGCCGAGTCGGGTCGCCCTGCGCAGACGACG TAACGCGGGACGGCGGCTGCTAGCCGTAGAGCGTCTGGCCTCCGAAGGCGACCGCCGCAGGAAGGGATTA CCTGCAGGGAAAGCGCGGTCCCGACGGGAGACGAGGCCGTGTGGACGGGCGGCAAAGCGTTGCCCGCTGG CCCGCGTACGTGTAGCCGAACTGCTGCGAGAAACGCAGAGACGGGAGCGCCCTGTCCGGGCACGCCCGCG< Amblyomma Americanum IST2b F 1; DNA; Amblyomma americanum>ACGTTGGATG AAGCCCGCGCTCCAAGC< Amblyomma Americanum IST2c R 1; DNA; Amblyomma americanum>ACGTTGGATG GCAGCAGTTCGGCTACACGTA< Amblyomma caj ennense amp 1; DNA; Amblyomma caj ennense> GAAGGGAGATCTCGCGCGGGGTGCGGGGAGGCGACGGGCGCGAAAGCGTTGTGCTCAGCA CAGTTCGGCCAGGCGCAGAGGCGCGCCGTGTGCGCGGTGACGAAGGGTTCCCCCGGACCC TGCGAAAAGGAACCGCCTCCTCGAGTGGGCGCGAAGTCGAGTACGGGTCGAACGGACGCG< Amblyomma caj ennense 1ST F 1; DNA; Amblyomma caj ennense>ACGTTGGATG CGAATGCGGTGTGCGCAGCACAGT< Amblyomma caj ennense 1ST F 2; DNA; Amblyomma caj ennense>ACGTTGGATG CGAAAGCGTTGTGCTCAGCACAGT< Amblyomma caj ennense 1ST R 1; DNA; Amblyomma caj ennense>ACGTTGGATG ACTTCGCGCCCACTCGAGGAG< Amblyomma caj ennense ITS ext 1; DNA; Amblyomma caj ennense> TCCCTTTCCGCAGGGTCC - - - < Amblyomma maculatum Actin F 1; DNA; Amblyomma maculatum>ACGTTGGATG GCCCTGGACTTCGAGCAG< Amblyomma maculatum Actin R 1; DNA; Amblyomma maculatum>ACGTTGGATG CCCGTCAGGAAGTTCGTAGG< Amblyomma maculatum Actin ext 1; DNA; Amblyomma maculatum> ACCGCCGCCTCGTCCT ~ ~< Amblyomma maculatum amp 1; DNA; Amblyomma maculatum> CCGCGCGAGAAGAAGGGAGATCTCGCGCCGGGTTGTGCGGGGAAACGACCGGGTGTGAACGCGTTGACGC TAGAGGAGAGTTTGTAGGCGCAGAGGCGCGCGCTGCCGTGCGGCGCGCTAAAGGGATCCTCACGGGCTCC GAGGGAAAGGAGTCTCTCTGCTCAAGCATTGTCTAGTACGGGTCGAACGGACGCGTATCTCGTTACGAGC GTTGTCCGCCGTCGGTCTAAGTGCTTCGCAGTCTCTGTCCCGAAAAAAACTGGGCCACTCCAGTTGGGGC AGGGGCGACGCAAAATCTTTTGTGAAGATGCCCCTACGCCAGGTTGTACGTTCGTCTCTCGCTCGAGGGA< Amblyomma maculatum ITS ext 1; DNA; Amblyomma maculatum> ACAATGCTTGAGCAGA - - - < Amblyomma maculatum ITS2 F 1; DNA; Amblyomma maculatum> ACGTTGGATG TTGTGCGGGAAACGACCGGGTGT< Amblyomma maculatum ITS2 R 1; DNA; Amblyomma maculatum> ACGTTGGATG AACGCTCGTAACGAGATACGCG< Dermacentor variabilis ext 1; DNA; Dermacentor variabilis> AGAAGGGCGTGCCCCGAAAGC< Dermacentor variabilis amp 1; DNA; Dermacentor variabilis> GGAGTACGTCGAGCTCCAGAGCTGGTCGCTCGTTCGTGTCACCGCAGCTGTGTGGGCACCCCTTCCGGGC TTCGTCGCAGGAATCTGAAGATTCTTTGCGAGGAGCGGGGAGGAGAAGGGCGTGCCCCGAAAGCGGTTCG ACGCGATAGCGCCGTCTGCGAGCGAGAAGAGCACGGCACGGCGCAGAAATTGCCGCGAAACGGAAAATGT CTCCTTCGAGAGCGTTGGCCGAGCTGACGCGTTCCGTCGTAGTCCGCCGTCGGTCCAAGTGCTTCGCAGT CTCTGTCCCCTAAAGACTGGGCCACTCCAGTTGGGGCAGGGGCGACGCTACACGAGACGATGCCCCCCGC< Dermacentor variabilis IST2b F 1; DNA; Dermacentor variabilis> ACGTTGGATG CTGAAGATTCTTTGCGAGGAGCGG< Dermacentor variabilis IST2b R 1; DNA; Dermacentor variabilis> ACGTTGGATG GCGTCAGCTCGGCCAAC< Dermacentor IST2 F 1; DNA; Dermacentor v a?: iabills>ACGTTGGATG GTGCGTCCGTCGACTCGTT< Dermacentor IST2 R 1; DNA; Dermacentor vaxiabilis>ACGTTGGATG TCGCCCAACACGGAGCTACT< Haemaphysalis longicornis ext 1; DNA; Haemaphysalis longicornis> aaTCCCGCTGAGCCCGCACTT< Haemaphysalis longicornis amp 1; DNA; Haemaphysalis longicornis> CGTACGAAAAAGGGCGAGTGTGCAGTTGCGGTCGTGGCTTTGCAGTAGACGGTTCTCTTTTGGGATGGAT GCTGTGATGGCTTTCCCGCTGAGCCCGCACTTGTAAGTGTGGGGCTCGCGGTACAACGAGAGTTGTCGCA TCCGGAGGCCGTTTTGCCTGTAAGCTTCCGCGCTTGACCGGGAACTTTTCCAGGTCGCGTTGGATGAAGC< Haemaphysalis longicornis ITS F 1; DNA; Haemaphysalis longicornis> ACGTTGGATG TCTTTTGGGATGGATGCTGTGATG< Haemaphysalis longicornis ITS R 1; DNA; Haemaphysalis longicornis> ACGTTGGATG CGGATGCGACAACTCTCGTTG- < Ixodes pacificus ext 1; DNA; Ixodes pacificus> AAGACCTTTGGGGACGGATGATGG< Ixodes pacificus amp 1; DNA; Ixodes pacificus> GTCCTCTTCCTACCCGGTTTGTGTCGTCGAACACTGCATAGAAAAACGAAACTCGATGGCGACCGTTTTG TGGAAAATCCCGAAACTTTCGCACGTTGAACGGCGCTTGTTGACGTCGGTGCGTTGGAAAAACGGAGATT TGAAACGGTTTCTTTTCGATCGATTCTGTTTTTCTTTTGGGCGTGGATGTCGTTCGAAAGACCTTTGGGG ACGGATGATGGACGGAGTTGGTTGGGCGTTGAAACGCGGCGTCTGCAATGAATCTTGTGGCGTTGATTTT GCAACACACTTCTTGCTGGGGAGAGAACGAGTACGTTTCTGGGAGGAAAGTGACTTTTTTTGCGTCGTAG CCTTCCGTCAGTCTAAGACCTTCGCGTCCCCGATGAATACTGGAGCCATCCAGTAGGGGTATGCCGTTGG< Ixodes pacificus IST2 F 1; DNA; Ixodes pacificus>ACGTTGGATG CTTCCTACCCGGTTTGTGTCGTC< Ixodes pacificus IST2 R 1; DNA; Ixodes pacificus>ACGTTGGATG CGTACTCGTTCTCTCCCCAGC< Ixodes scapularis ext 1; DNA; Ixodes scapularis> TGCGCTTAACCAGTCCTCCTCCT< Ixodes scapularis amp 1; DNA; Ixodes scapularis> CTGTGTGAGGGTCGGATCATATATCAAGAGAGGAGAATTTGTTTTCTACCTCGTTTTGACTGTGTCGGAT CGTGGGCAGCACGCAGTTTTATGCTTTCTTGCGTTGCGTTTTCTTTGAGCAAATGCACGAGTGGTGCGAT TGCACGCGTGCGCTTAACCAGTCCTCCTCCTCCTACGAGTTTTTATCGAACACTGCATGGGAAAACGAAA CTCGATGGATACCGTTTGTGGAAAATCCCGTACCAAAAAAATCTTTCGCACGTTGAACGGCGCTGTGACG TCGGTGCGTTAGAAACGGAGATTTGAAACGGTTTCTTTTCGATCGATTCTGTTTTCTTTGGCGTGGATGT< Ixodes scapularis F 1; DNA; Ixodes scapularis>ACGTTGGATG TGCGTTTTCTTTGAGCAAATGCACGAG< Ixodes scapularis R 1; DNA; Ixodes scapularis>ACGTTGGATG GTACGGGATTTTCCACAAACGGTATCCA< Tick spp F 1; DNA; Ixodes sp. >ACGTTGGATG CTGCTCAATGATTTTTTAAATTGCTGTGGT< Tick spp R 1; DNA; Ixodes sp. >ACGTTGGATG CCGGTCTGAACTCAGATCAAGTAGGAHOST IDENTIFICATION (BLOOD MEAL) SEQUENCES (SEQ ID NOS: 419-569)< Bear (brown, black bear) ext 1; DNA; Ursus americanus>TGC CTA CGC TAT CCT ACG ATC< Bear (brown, black bear) Amp 1; DNA; Ursus americanus> ACTATACAATTAAGGATATTCTAGGCGCCCTACTTCTCACCCTAGCCTTAGCAACCCTAGTCCTATTCTC G CCCGACTTACTAGGAGACCCTGACAAC T AT AT C C C C G C AAAT C C AC T GAG C AC C C C AC C C C AC AT C AAA CCCGAGTGGTACTTTCTATTTGCCTACGCTATCCTACGATCCATCCCTAATAAACTAGGAGGAGTACTAG CACTAATTTTCTCCATTCTAATCCTAGCCCTCATTCCTCTTCTACACACGTCCAAACAACGAGGAATGAT ATTCCGGCCCCTAAGCCAATGCCTATTTTGACTTCTAGTAGCAGACCTACTAACACTAACATGAATTGGA GGACAACCAGTAGAACACCCCTTCATTATTATCGGACAACTAGCCTCCATTCTCTACTTTACAATCCTCC TAGTACTTATACCCATCGCTGGAATTATTGAAAACAACCTCTTAAAGTGGAGAGTCTTTGTAGTATAACA< Bear (brown, black bear) F 1; DNA; Ursus americanus>ACGTTGGATG CCGACTTACTAGGAGACCCTG< Bear (brown, black bear) R 1; DNA; Ursus americanus>ACGTTGGATG GTCAAAATAGGCATTGGCTTAGG< Bird-l. l Ext 1; DNA; Aves>ttTgccaggCtggatggg< Bird-1. 2 Ext 2; DNA; Aves>ttCgccaggTtggatggg<birdl E l l; DNA; Aves>Cccagagaagttgtgg<birdl F 1; DNA; Aves>ACGTTGGATG cccagagaagttgtgg<birdl F 2; DNA; Aves>ACGTTGGATG cccagagaagctgtgg<birdl F 2 1; DNA; Aves>Cccagagaagctgtgg<birdl R 1; DNA; Aves>ACGTTGGATG catgggcagggacacc<birdl R 1 1; DNA; Aves>Catgggcagggacacc<bird-2. 1 Ext 1; DNA; Aves>atCgcccagagaagctgtgg<bird-2. 2 Ext 2; DNA; Aves>ataGCCCAGAGCAGCTGTGG<bird2 F 1; DNA; Aves>ACGTTGGATG SAGGCCCTGGCACAGG<bird2 R 1; DNA; Aves>ACGTTGGATG CCTTGRACACTTCCAGGGAT< Bobcat-Lynx Ext 1; DNA; Lynx rufus>CCT TGA ATT CGC TGT GGC< Bobcat-lynx F 1; DNA; Lynx rufus>ACGTTGGATG GCAAACATCAGCACCTCCGTT< Bobcat-lynx R 1; DNA; Lynx rufus>ACGTTGGATG CTAGTAGGGTGAAGACGTAGGCTTG<cat F 1; DNA; Felis catus>ACGTTGGATG CTTCGGATTCTGTGTCTCC<cat R 1; DNA; Felis catus>AC GT T GGAT G TAT T T T T GGGACAGAGAGAGAC<cat Ext 1; DNA; Felis catus>attcTCTGaCCCTCCCCCGT<cat Ext 2; DNA; Felis catus>attaTCTGcCCCTCCCCCGT<deer Ext 1; DNA; Odocoileus virginianus> CGATGCTGTTCTCTTGAAACATCCC<deer 0 F 1; DNA; Odocoileus virginianus>ACGTTGGATG GATCTGTTTCACCCTAGATAAT<deer 0 R 1; DNA; Odocoileus virginianus>ACGTTGGATG CTGTGGGAGAAGGCGAG<deer 1 F 1; DNA; Odocoileus virginianus>GATCTGTTTCACCCTAGATAAT<deer 1 R 1; DNA; Odocoileus virginianus>CTGTGGGAGAAGGCGAG<deer mouse ext 1; DNA; Peromyscus arboreus>CATCTGCCGAGACGT- < Dog / canis lupis Amp 1; DNA; Canis sp. >T GAAAT T GACCTTCCCGTGAAGAGGCGGGAATACCACAATAAGAC GAGAAGAC CCTATGGAGCTT T AAT T AACTAACCCAAACTTATGGATACTAGATACCTACAAGGCATAACATAACACCATTATTATGGGTTAGCAA TTTAGGTTGGGGTGACCTCGGAATATAAAAAAACTC C C GAGT GAT T AAAAT T T AGAC C CACAAGT CAAAA TACAACATCACTTATTGATCCAATAATTTTTGATCAACGGAACAAGTTACCCTAGGGATAACAGCGCAAT< Dog / canis lupis ext 1; DNA; Canis sp. >CCCAAACTTATGGATACTAG- < Dog / canis lupis F 1; DNA; Canis sp. >CTTCCCGTGAAGAGGCGGGAATAC< Dog / canis lupis R 1; DNA; Canis sp. >TTTAGGTTGGGGTGACCTCGGA<felid F 1; DNA; Felis sp. >AC GT T G GAT G CCTATTTAACCTACCACACC C AC AAG<felid R 1; DNA; Felis sp. >ACGTTGGATG GCCAGATGCTTTGTTTAAGCTACATC<felids R 1 1; DNA; Felinae>GCCAGATGCTTTGTTTAAGCTACATC<felids ext 1; DNA; Felis sp. >G GAGACAAGT C GT AACAAGG<felids F 1; DNA; Felis sp. >CCTATTTAACCTACCACACC C AC AAG<felids R 1; DNA; Felis sp. >GCCAGATGCTTTGTTTAAGCTACATC<felids F 1 1; DNA; Felinae>C C T AT T T AAC CT AC C AC AC C C AC AAG< Fox / Vulpes extl; DNA; Vulpes vulpes>C AAAC C CAT GAAAT C C AAAC C C C< Fox / Vulpes amp 1; DNA; Vulpes vulpes> CATTTGTTCCTTAAATAGGGACTTGTATGAATGGCCACACGAGGGTTTAACTGTCTCTTACTTCCAATCC GTGAAATTGACCTTCCCGTGAAGAGGCGGGAATATCATAATAAGACGAGAAGACCCTATGGAGCTTTAAT TAATTAGCCCAAACCCATGAAATCCAAACCCCTCCGGGAATAACTTACTATCATCGTTATGGGCTAACAA TTTAGGTTGGGGTGACCTCGGAATATAAAAAAAC T C C C GAGT GAT T AAAAT TTAGACCTACCAGT CAAAA TGTATCATCACTTATTGATCCAATAATCATTGATCAACGGAACAAGTTACCCTAGGGATAACAGCGCAAT< Fox / Vulpes ext 2; DNA; Vulpes vulpes>C AAAC C CAT GAAAT C C AAAC C C C< Fox / Vulpes F 1; DNA; Vulpes vulpes>CTTCCCGTGAAGAGGCGGGAATA< Fox / Vulpes R 1; DNA; Vulpes vulpes>TTTAGGTTGGGGTGACCTCGGA< Fox Dog F 1; DNA; Vulpes vulpes>ACGTTGGATG CTTCCCGTGAAGAGGCGGGAATAC< Fox Dog R 1; DNA; Vulpes vulpes>ACGTTGGATG- TCCGAGGTCACCCCAACCTAAA< Goat ext 1; DNA; Capra hircus>CAG GCA TTC ACC CAC TTA ATCC< Goat (Capra aegagrus hircus ) amp 1; DNA; Capra hircus> ATTTCCGAAGTTTTATTCTTTACTGGGTTTTTCTGAGCTTTCTATCACTCGAGCCTTGCCCCCACACCCG AATTAGGCGGCTGCTGACCTCCAACAGGCATTCACCCACTTAATCCCCTAGAAGTCCCATTACTTAATAC TTCCGTCCTCCTAGCCTCAGGAGTTTCCATCACCTGAGCTCACCATAGCCTTATGGAAGGAGACCGTAAC CACATACTACAAGCCTTATTCATTACCATTATACTAGGCTTATACTTCACATTATTACAAGCATCAGAAT< Goat F 1; DNA; Capra hircus>ACGTTGGATG GGTTTTTCTGAGCTTTCTATCACTCG< Goat R 1; DNA; Capra hircus>ACGTTGGATG TAAGCCTAGTATAATGGTAATGAATAAGG< Goose Anser anser amp 1; DNA; Anser anser> AACAGTGGACACAATAGCACCCCGCTAATAAGACAGGTCAAGGTATAGCCTATGGAGTGGAAGAAATGGGCTACATT CCCTATTCATAGGGCACACGGAAAGAAGCGTGAAACCACTTCTGGAAGGCGGATTTAGCAGTAAAGTGGGATAATAG AGCCTACTTTAAGCCGGCCCTGGGGC< Goose Anser anser ext 1; DNA; Anser anser>GCACACGGAAAGAAGCGT”< Goose Anser anser F 1; DNA; Anser anser>TCAAGGTATAGCCTATGGAGTGA< Goose Anser anser R 1; DNA; Anser anser>CATTCTGGAAGGCGGATTTAG< Goose Anser cygnoides amp 1; DNA; Anser cygnoides> AACAGTGGACACAATAGCACCCCGCTAATAAGACAGGTCAAGGTATAGCCTATGGAGTGGAAGAAATGGGCTACATT CCCTATTCATAGGGCACACGGAAAGAAGCGTGAAACCACTTCTGGAAGGCGGATTTAGCAGTAAAGTGGGACAATAG AGCCTACTTTAAGCCGGCCCTGGGGC< Goose Anser cygnoides ext 1; DNA; Anser cygnoides>GCACACGGAAAGAAGCGT< Goose Anser cygnoides F 1; DNA; Anser cygnoides>TCAAGGTATAGCCTATGGAGTGA< Goose Anser cygnoides R 1; DNA; Anser cygnoides>CATTCTGGAAGGCGGATTTAG< Goose Anser fabalis amp 1; DNA; Anser fabalis> AACAGTGGACACAATAGCACCCCGCTAATAAGACAGGTCAAGGTATAGCCTATGGAGTGGAAGAAATGGGCTACATT CCCTATTCATAGGGCACACGGAAAGAAGCGTGAAACCACTTCTGGAAGGCGGATTTAGCAGTAAAGTGGGACAATAG AGCCTACTTTAAGCCGGCCCTGGGGC< Goose Anser fabalis ext 1; DNA; Anser fabalis>GCACACGGAAAGAAGCGT< Goose Anser fabalis F 1; DNA; Anser fabalis>TCAAGGTATAGCCTATGGAGTGA< Goose Anser fabalis R 1; DNA; Anser fabalis>CATTCTGGAAGGCGGATTTAG< Goose Branta nigricans amp 1; DNA; Branta sp. > AACAGTGGACACAATAGCACCCCGCTAATAAGACAGGTCAAGGTATAGCCTATGGGGTGGAAGAAATGGGCTACATT CCCTATTCATAGGGCACACGGAAAGAAGCGTGAAACCGCTTCTGGAAGGCGGATTTAGCAGTAAAGTGGGACAATAG AGCCTACTTTAAACCGGCCCTGGGGC< Goose Branta nigricans ext 1; DNA; Branta sp. >GCACACGGAAAGAAGCGT< Goose Branta nigricans F 1; DNA; Branta sp. >TCAAGGTATAGCCTATGGAGTGA< Goose Branta nigricans R 1; DNA; Branta sp. >CATTCTGGAAGGCGGATTTAG< Goose 0 ext 1; DNA; Anser anser>GCACACGGAAAGAAGCGT< Goose 0 F 1; DNA; Anser anser>ACGTT GGAT G T CAAGGTATAGCCTAT GGR GT GGA< Goose Anser anser R 1; DNA; Anser anser>ACGTTGGATG CTAAATCCGCCTTCCAGAAGYG< Horse 0 ext 1; DNA; Equus caballus>tatTACACCAGTCTCAGCCCTACT< Horse 0 F 1; DNA; Equus caballus>ACGTTGGATG- ATCTTCATACTCGACCCCAACC< Horse 0 R 1; DNA; Equus caballus>ACGTTGGATG- AGCGGATTAGCAGGAAGACG< Horse Equus caballus amp 1; DNA; Equus caballus> GCTTCATCATGGCCATAGCCTGATTCCTATTCAACACCAACACATGAGACCTCCAACAAATCTTCATACT CGACCCCAACCTTACCAACCTCCCGCTCCTAGGCCTCCTCCTAGCCGCAACTGGCAAATCCGCTCAATTT GGACTCCACCCATGACTTCCTTCAGCCATAGAGGGCCCTACACCAGTCTCAGCCCTACTCCACTCCAGCA C AAT AGT T GT AG C AG GCGTCTTCCTGCTAATCCGCTT C CAT C C AC T AAT AGAAAAC AAC AAAAC AAT C C A GTCACTTACCCTATGCCTAGGAGCCATCACCACACTATTCACAGCAATCTGCGCACTCACTCAAAACGAT< Horse Equus caballus ext 1; DNA; Equus caballus> CTACACCAGTCTCAGCCCTACTCCAC< Horse Equus caballus F 1; DNA; Equus caballus>CGACCCCAACC< Horse Equus caballus R 1; DNA; Equus caballus> CGTCTTCCTGCTAATCCGCT< Hu RNaseP ext 1; DNA; homo sapiens> ACCCTGGGCGTCAATATGG< Hu RNaseP ext 2; DNA; homo sapiens> TTCTGACCTGAAGGCTCTGCGC< Hu RNaseP F 1; DNA; homo sapiens>ACGTTGGATG- AGATTTGGACCTGCGAGCG< Hu RNaseP R 1; DNA; homo sapiens>ACGTTGGATG” GAGCGGCTGTCTCCACAAGT< Hu (i2 megaglobulin ext 1; DNA; homo sapiens>C CAT GTGACTTTGTCACAGCC C AAG< Hu (i2 megaglobulin F 1; DNA; homo sapiens> ACGTTGGATG TGAGTATGCCTGCCGTGTGA< Hu (i2 megaglobulin R 1; DNA; homo sapiens> ACGTTGGATG ACTCATACACAACTTTCAGCAGCTTAC< Hu GAPDH ext 1; DNA; Homo sapiens> AGAGCTAGGAAGGACAGGC< Hu GAPDH F 1; DNA; Homo sapiens>ACGTTGGATG CTCCCCACACACATGCACTTA< Hu GAPDH R 1; DNA; Homo sapiens>ACGTTGGATG CCTAGTCCCAGGGCTTTGATT< Mouse 0 Ext 1; DNA; Mus mus culus> TGGGCTACCAAGTGAGCTCCAG< Mouse 0 F 1; DNA; Mus mus culus>ACGTTGGATG- GATCTCTGTGAGTTCGAGG< Mouse 0 R 1; DNA; Mus mus culus>ACGTTGGATG- GTTTCTCTGTGTAGCTTTGC<mouse 1 F 1; DNA; Mus mus culus> GTTTCTCTGTGTAGCTTTGC<mouse 1 R 1; DNA; Mus mus culus> GATCTCTGTGAGTTCGAGG<mouse Sub F 1; DNA; Mus mus culus>ACGTTGGATG- AGCCATATGCACTACACATC<mouse Sub R 1; DNA; Mus mus culus>ACGTTGGATG- GAG GCT CCG TTT GCG TGT A< Pig 0 ext 1; DNA; Sus sp. >GT ACA- TAG- TCT CCT CAT TAG CC< Pig_0_F_l; DNA; Sus sp. >ACGTTGGATG ACA ACA TAA TCT GAA TCA ATG C< Pig_0_R_l; DNA; Sus sp. >ACGTTGGATG TTC GCC TAG TTG GTT TAG TAG< Pig_Amp_l; DNA; Sus sp. >ACAACATAAT CT GAAT CAAT GCa a cagtacatagtctcctcattagcct g atcagtctatccCTACTAAACCAACTAGGCGAA< Pig ext 1; DNA; Sus sp. >gtacatagtctcctcattagcc< Pig_F_l; DNA; Sus sp. >ACAACATAAT CT GAAT CAAT GC< Pig_R_l; DNA; Sus sp. >TACTAAACCAACTAGGCGAA< Pig_R_2; DNA; Sus sp. >TACTAAACCAACTAGGCGAA<rabbit 0 ext 1; DNA; Oryctolagus cuniculus> attGGCCCTGCACCCCATG<rabbit 0 F 1; DNA; Oryctolagus cuniculus> ACGTTGGATG ggaaggcagtggaggat<rabbit 0 F 1; DNA; Oryctolagus cuniculus> ACGTTGGATG ggtgcttcctcctggtct<rabbit F 1; DNA; Oryctolagus cuniculus>Ggaaggcagtggaggat<rabbit R 1; DNA; Oryctolagus cuniculus>Ggtgcttcctcctggtct< Raccoon ext 1; DNA; Procyon lotor>CCCAAATTTATGGCCA< Raccoon-dog Ext 1; DNA; Procyon lotor>CAGGTCTATTTCTAGCC< Raccoon-dog Amp 1; DNA; Procyon lotor> ATCGCACCCACTAGCCAAAATCGTTAACAACTCATTTATTGATCTCCCCGCACCATCAAACATCTCTGCC TGATGAAACTTCGGATCACTGCTAGGAGTATGTCTTATTCTACAGATTATAACAGGTCTATTTCTAGCCA TAG AC TAT AC AT CAGATACCTCTACTGCCTTTTCATCAGT TAG AC AT AT CTGCCGAGACGT C AAT T AC G G< Raccoon-dog F 1; DNA; Procyon lotor>ACGTTGGATG CTGATGAAACTTCGGATCACTG< Raccoon-dog F 2; DNA; Procyon lotor>C TGATGAAACTTCGGATCACTG< Raccoon-dog R 1; DNA; Procyon lotor>ACGTT GGAT G CT GAT GAAAAGGCAGTAGAGGTA< Raccoon-dog R 2; DNA; Procyon lotor>TACCTCTACTGCCTTTTCATCAG< Sheep (Ovis aries ) amp 1; DNA; Ovis aries>AAACAT GAAACAT CGGAGTAATCCTCCTATTTGCGACAATAGCCACAGCATTCATAGGCTAT GT T T TAG C ATGAGGACAAATATCATTCTGAGGAGCAACAGTTATTACCAACCTCCTTTCAGCAATTCCATATATTGGC ACAAACCTAGTCGAATGAATCTGAGGAGGATTCTCAGTAGACAAAGCTACCCTCACCCGATTTTTCGCCT TTCACTTTATTTTCCCATTCATCATCGCAGCCCTCGCCATAGTTCACCTACTCTTCCTCCACGAAACAGG< Sheep (Ovis aries ) ext 1; DNA; Ovis aries>AGCCACAGCATTCATAGGCT< Sheep (Ovis aries ) F 1; DNA; Ovis aries>GGAGTAATCCTCCTATTTGCG< Sheep (Ovis aries ) R 1; DNA; Ovis aries>TTTCCCATTCATCATCGC< Sheep 0 ext 1; DNA; Ovis aries>ACAGCATTCATAGGCT< Sheep 0 F 1; DNA; Ovis aries>ACGTTGGATG- GGAGTAATCCTCCTATTTGCG< Sheep 0 R 1; DNA; Ovis aries>ACGTTGGATG- GCGATGATGAATGGGAAA<shrew Ext 1; DNA; Sorex cinereus>attTCCCCCAAGCACCGCCAG<shrew F 1; DNA; Sorex cinereus>ACGTTGGATG GATTCCCAGCATCCCATATG<shrew F 2; DNA; Sorex cinereus>GAT T C C C AG CAT C C CAT AT G<shrew R 1; DNA; Sorex cinereus>ACGTTGGATG RTTACTCCTGGCTCTGCA<shrew R 2; DNA; Sorex cinereus>RTTACTCCTGGCTCTGCA<squirrel Ext 1; DNA; Sciurus carolinensis>GGGCTAGGGATGTGGCTCAGTG<squirrel F 1; DNA; Sciurus carolinensis>ACGTTGGATG ccctgtctct aaataaaata ca<squirrel F 2; DNA; Sciurus carolinensis>ccctgtctct aaataaaata ca<squirrel R 1; DNA; Sciurus carolinensis>ACGTTGGATG TACCAGGGATTGAACTCAG<squirrel R 2; DNA; Sciurus carolinensis>TACCAGGGATTGAACTCAG< Turkey ext 1; DNA; Meleagris gallopavo>ccgccAACGTTTCGTTT< Turkey mitochondrial partial D-loop amp 1; DNA; Meleagris gallopavo> CGGACATATTTTTGCTAATTTTCACTTCCTCTATTTTCCCAACAAAACCAGGAAATTACCCACATTTTTTCCCCGCT AAAAC C C CACAAACAAT AAAAACGTTTCGTTTAAT AT AT ACAT AT AT T T T T T T T T GCT AT T T T TAT T AGAGAAAC C CCCCTACCAAACCTATCATTCCTAAAAACAAAAATTGCACAGCACAAAACTCCACGAACAAACATTATTTATATTAAC AATTAACAAACAGCTAAATTCTCCTACTACCAACA <Turkey ext 2; DNA; Meleagris gallopavo>AACGTTTCGTTTA <Turkey F 1; DNA; Meleagris gallopavo>ACGTTGGATG GCATAAAACCCCACAAACAATAA <Turkey F 2; DNA; Meleagris gallopavo>GCTAAAACCCCACAAACAATAA <Turkey R 1; DNA; Meleagris gallopavo>ACGTTGGATG GGGTTTCTCTAATAAAAATAGCAAAAA <Turkey R 2; DNA; Meleagris gallopavo>T T T T T G C T AT T T T T AT T AGAGAAAC C C<vole Ext 1; DNA; Microtus arvalis>ttCTCTTCTGGCCTGCAG<vole F 1; DNA; Microtus arvalis>ACGTTGGATG TGAGTTCAATTCCCAGCAAC<vole F 2; DNA; Microtus arvalis>T GAGT T CAAT T C C CAGCAAC<vole R 1; DNA; Microtus arvalis>AC GT T GGAT G T GT AT ACAAT AT TCTGTCTGTGTG<vole R 2; DNA; Microtus arvalis>ACGTT GGAT G T GTATACAATATT CT GT CT GCAT G<vole R 3; DNA; Microtus arvalis>T GTATACAATATT CT GT CT GT GT G<vole R 4; DNA; Microtus arvalis>T GTATACAATATT CT GT CT GCAT G<white foot mouse ext 1; DNA; Peromyscus leucopus>cACAGCATTCTCATCCG<Wild boar Ext 1; DNA; Sus scrofa>C T CACAGGT GAT GT GAC<Wild boar F 1; DNA; Sus scrofa>ACGTTGGATG” GTAAGAAAATCTTAACCTAGCAAATGGGT<Wild boar F 2; DNA; Sus scrofa>GTAAGAAAATCTTAACCTAGCAAATGGGT<Wild boar R 1; DNA; Sus scrofa>ACGTTGGATG- AGGGAGTTTTTTGTTCTTACCCG<Wild boar R 2; DNA; Sus scrofa>AGGGAGTTTTTTGTTCTTACCCGCLAIMSWhat is claimed is:1. A method of identifying (i) an arthropod-borne pathogen, (ii) an arthropod species, and (iii) a blood meal host from a single biological sample, the method comprising:(a) amplifying nucleic acids in the biological sample with a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three arthropod species, and (iii) PCR primers specific for at least three blood meal hosts;(b) performing single base extension with a set of extension primers on the amplified products to generate a set of extended products; and(c) analyzing the set of extended products so as to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).2. The method of claim 1, wherein the arthropod-borne pathogen is a tick-borne pathogen and the arthropod species is a tick.3. The method of claim 1, wherein the arthropod-borne pathogen is a mosquito-borne pathogen and the arthropod species is a mosquito.4. The method of claim 1, wherein the arthropod species is a biting midge, a sandfly, or a tsetse fly.5. The method of claim 1, wherein the arthropod species is a flea, a louse, or a triatomine bug.6. The method of claim 1, wherein the set of PCR primers comprise (i) PCR primers specific for at least fifteen arthropod-borne pathogens.7. The method of claim 1, wherein the set of PCR primers comprise (i) PCR primers specific for at least twenty arthropod-borne pathogens.

Claims

8. The method of claim 1, wherein the set of PCR primers comprise (ii) PCR primers specific for at least five species of arthropods.

9. The method of claim 1, wherein the set of PCR primers comprise (iii) PCR primers specific for at least ten blood meal hosts.

10. The method of claim 1, wherein the arthropod-borne pathogen is a bacterium, a fungus, a protozoan, or a virus.

11. The method of claim 10, wherein the arthropod-borne pathogen is a bacterium and the PCR primers specific for at least ten arthropod-borne pathogens hybridize with a 16S and / or an 18S ribosomal nucleic acid of the bacterium.

12. The method of claim 10 wherein the arthropod-borne pathogen is a bacterium and the bacterium is an Anaplasma, a Bartonella, a Borrelia, a Coxiella, an Ehrlichia, a Francisella, a Rickettsia, or a Yersinia.

13. The method of claim 12, wherein the Anaplasma, the Babesia, the Bartonella, the Borrelia, the Coxiella, the Ehrlichia, the Francisella, the Rickettsia, or the Yersinia are Anaplasma bovis, Anaplasma caudatum, Anaplasma centrale, Anaplasma marginale, Anaplasma mesaenterum, Anaplasma odocoilei, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma platys, Bartonella bacilliformis, Bartonella elizabethae, Bartonella henselae, Bartonella quintana, Bartonella vinsonii, Borrelia afzelii, Borrelia americana, Borrelia anserina, Borrelia bissettiae, Borrelia burgdorferi, Borrelia californiensis, Borrelia carolinensis, Borrelia coriaceae, Borrelia garinii, Borrelia hermsii, Borrelia kurtenbachii, Borrelia lanei, Borrelia lonestari, Borrelia lusitaniae, Borrelia mayonii, Borrelia miyamotoi, Borrelia parkeri, Borrelia theileri, Borrelia turicatae, Borrelia valaisiana, Coxiella burnetii, Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia mineirensis, Ehrlichia muris, Ehrlichia muris eauclairensis, Ehrlichia muris-like agent, Ehrlichia ruminantium, Ehrlichia sp., Francisella tularensis,Panola Mountain Ehrlichia, Rickettsia africae (aeschlimannii ), Rickettsia akari, Rickettsia amblyommatis, Rickettsia australis, Rickettsia bellii, Rickettsia CA6269, Rickettsia conorii, Rickettsia felis, Rickettsia heilongjiangensis, Rickettsia helvitica, Rickettsia honei, Rickettsia japonica, Rickettsia massiliae, Rickettsia monacensis, Rickettsia montanesis, Rickettsia parkeri, Rickettsia philippi (strain 364D), Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia sibirica, Rickettsia slovaca, Rickettsia typhi, or Yersinia pestis.

14. The method of claim 10, wherein the arthropod-borne pathogen is a protozoan, and the protozoan is a Babesia, a Cytauxzoon, a Leishmania, Plasmodium, a Theileria, or a Trypanosome.

15. The method of claim 14, wherein the Babesia is Babesia beliceri, Babesia bigemina, Babesia bovis, Babesia caballi, Babesia canis, Babesia catia, Babesia conradae, Babesia crassa, Babesia divergens, Babesia duncani, Babesia felis, Babesia foliatad, Babesia taylori, Babesia gibsoni, Babesia hongkongensis, Babesia jakimovi, Babesia lengau, Babesia major, Babesia microti, Babesia motasi, Babesia occultans, Babesia orientalis, Babesia ovata, Babesia ovis, Babesia perroncitoi, Babesia presentii, Babesia rossi, Babesia trautmanni, Babesia Venator um, or Babesia vogeli.

16. The method of claim 14, wherein the Cytauxzoon is Cytauxzoon banethi, Cytauxzoon europaeus, Cytauxzoon felis. Cytauxzoon manul or Cytauxzoon otrantorum.

17. The method of claim 14, wherein the Leishmania is Leishmania aethiopica, Leishmania amazonensis, Leishmania archibaldi, Leishmania braziliensis, Leishmania colombiensis, Leishmania donovani, Leishmania garnhami, Leishmania guyanensis, Leishmania infantum, Leishmania killicki, Leishmania lainsoni, Leishmania lindenbergi, Leishmania major, Leishmania mexicana, Leishmania naifft, Leishmania panamensis, Leishmaniaperuviana, Leishmania pifanoi, Leishmania shaw i, Leishmania tropica, or Leishmania venezuelensis.

18. The method of claim 14, wherein the Plasmodium is Plasmodium falciparum, Plasmodium know le si, Plasmodium malar iae, Plasmodium ovale curtisi, Plasmodium ovale wallikeri, Plasmodium sp., o Plasmodium vivax.

19. The method of claim 14, wherein the Theileria is Theileria annae, Theileria annulata, Theileria buffeli, Theileria equi, Theileria haneyi, Theileria lestoquardi, Theileria luwenshuni, Theileria mutans, Theileria orientalis, Theileria ovis, Theileria parva, Theileria separata, Theileria sergenti, Theileria taurotragi, Theileria uilenbergi, or Theileria velifera.

20. The method of claim 14, wherein the Trypanosome is Trypanosoma brucei, Trypanosoma congolense, Trypanosoma cruzi, Trypanosoma evansi, Trypanosoma equiperdum, Trypanosoma rangeli, Trypanosoma simiae, or Trypanosoma vivax.

21. The method of claim 10, wherein the arthropod-borne pathogen is a virus and the virus is African horse sickness virus, African swine fever virus, Aino virus (AINV), Akabane virus (AKAV), Alkhurma hemorrhagic fever virus, Barmah forest virus (BFV), Bebaru virus (BENV), Bhanja virus, Bluetongue virus, Bourbon virus, Bovine ephemeral fever virus, Bunyamwera virus (BUNV), Cache Valley virus (CVV), Chandipura virus (CHPV), Chikungunya virus (CHIKV), Colorado tick fever virus, Crimean Congo hemorrhagic fever virus (CCHFV), Dengue virus (DENV), Eastern equine encephalitis virus (EEEV), Edge Hill virus, Equine encephelitis virus, Everglades virus (EVEV), Getah virus (GETV), Guaroa virus (GROV), Heartland virus, Ibaraki virus, Ilheus virus (ILHV), Jamestown Canyon virus (JCV), Japanese encephalitis virus (JEV), Kairi virus, Kasba virus, Kyasunur Forest disease virus (KFDV), La Crosse virus (LACV), Main drain virus (MDV), Mayaro virus (MAYV), Middelburg virus (MIDV), Murray Valleyencephalitis virus (MVEV), Nairobi sheep disease virus, Ndumu virus (NDUV), Omsk hemorrhagic fever virus, O'nyong nyong virus (ONNV), Oropouche virus (OROV), Peruvian horse sickness virus, Powassan virus (POWV), Rift Valley fever virus (RVFV), Ross River virus (RRV), Sandfly fever virus, Schmallenberg virus (SBV), Semliki forest virus (SFV), Shuni virus, Sindbis virus (SINV), Snowshoe hare virus (SSHV), St. Louis encephalitis virus (SLEV), Tembusu virus, Thogoto virus, Tick borne encephalitis virus (TBEV), Tyuleniy virus, UNA virus (UNAV), Usutu virus (USUV), Venezualan equine encephalitis virus (VEEV), Vesicular stomatitis virus (VSV), Wesselsbron virus, West Nile virus (WNE), Western equine encephalitis virus (WEEV), Yellow fever virus (YFV), Yunnan virus, or Zika virus (ZIKV).

22. The method of claim 1, wherein the PCR primers specific for at least three arthropod species hybridize with a gene from a ribosomal 12S subunit, a ribosomal 16S subunit, a ribosomal 18S subunit, or a cytochrome c oxidase unit 1 of the at least three arthropod species.

23. The method of claim 2, wherein the tick species comprise a soft tick or a hard tick.

24. The method of claim 23, wherein the tick species is Amblyomma americanum, Amblyomma cajennense, Amblyomma maculatum, Amblyomma sp., Amblyomma testudinarium, Amblyomma variegatum, Argas persicus, Bothriocroton hydrosauri, Dermacentor albipictus, Dermacentor andersoni, Dermacentor nitens, Dermacentor occidentalis, Dermacentor reticulatus, Dermacentor sp., Dermacentor variabilis, Haemaphysalis longicornis, Haemaphysalis sp., Hyalomma marinatum, Hyalomma sp., Hyalomma truncation, Ixodes cookei, Ixodes holocyclus, Ixodes pacificus, Ixodes persulcatus, Ixodes ricinus, Ixodes scapular is, Ixodes sp., Ixodes spinipalpis, Margaropus winthemi, Ornithodoros moubata, Ornithodoros rudis, Ornithodoros savignyi, Ornithodoros turicatae, Otobius megnini, Rhipicephalus annlatus,Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Rhipicephalus sanguineus, or Rhipicephalus sp.

25. The method of claim 3, wherein the mosquito is n Aedes, an Anopheles, a Coquillettidia, a Culex, a Culiseta, a Haemagogus, an Ochlerotatus, or a Psorophora mosquito.

26. The method of claim 25, whereinAedes, the Anopheles, the Coquillettidia, the Culex, the Culiseta, the Haemagogus, the Ochlerotatus, or the Psorophora mosquito is Aedes aegypti, Aedes albopictus, Aedes bahamensis, Aedes fulvus pallens, Aedes infirmatus, Aedes sollicitans, Aedes sp., Aedes tormentor, Aedes taeniorhynchus, Aedes triseriatus, Aedes vexans, Anopheles albimanus, Anopheles arabiensis, Anopheles atroparvus, Anopheles barbirostris, Anopheles christyi, Anopheles claviger, Anopheles coluzzii, Anopheles coustani, Anopheles culicifacies, Anopheles darlingi, Anopheles dims, Anopheles epiroticus, Anopheles farauti, Anopheles funestus, Anopheles gambiae, Anopheles maculatus, Anopheles melas, Anopheles merus, Anopheles minimus, Anopheles nimbus, Anopheles pseudopunctipennis, Anopheles quadriannulatus, Anopheles rufipes, Anopheles sacharovi, Anopheles sinensis, Anopheles sp., Anopheles stephensi, Anopheles walker, Coquillettidia perturbans, Coquillettidia sp., Coquillettidia venezuelensis, Culex biscaynensis, Culex declarator, Culex erraticus, Culex fatigans, Culex pedroi, Culex pipiens, Culex quinquefasciatus, Culex sp., Culiseta dyari, Culiseta inornata, Culiseta melanora, Culiseta sp., Haemagogus capricornii, Haemagogus equinus, Haemagogus janthinomys, Haemagogus lucifer, Haemagogus sp., Ochlerotatus albifasciatus, Ochlerotatus sp., Ochlerotatus triseriatus, Psorophora confmnis, Psorophora ferox, or Psorophora sp.

27. The method of claim 4, wherein the biting midge, the sandfly, or the tsetse fly are Culicoides sp., Glossina sp., Lutzomyia sp. or Phlebotomus sp.

28. The method of claim 27, wherein the Culicoides sp. biting midge is Culicoides actoni,Culicoides adersi. Culicoides brevitarsis, Culicoides fulvus, Culicoides furans, Culicoides grahamii, Culicoides imicola, Culicoides inornatipennis, Culicoides insignis, Culicoides insinuatus, Culicoides milnei, Culicoides obsoletus, Culicoides oxystoma, Culicoides paraensis, Culicoides phlebotomus, Culicoides schultzei, Culicoides variipennis, or Culicoides wadai.

29. The method of claim 27, wherein the Glossina sp. tsetse fly is Glossina austeni, Glossina brevipalpis, Glossina fusca, Glossina fuscipes, Glossina longipalpis, Glossina morsitans, Glossina palpalis, Glossina tabaniformis, Glossina tachinoides, or Glossina vanhoofi.

30. The method of claim 27, wherein the Lutzomyia sp. or Phlebotomus sp. sandfly is Lutzomyia amazonensis, Lutzomyia anduzei, Lutzomyia aracuchensis, Lutzomyia ayrozai, Lutzomyia carerrai, Lutzomyia christophei, Lutzomyia complexa, Lutzomyia diabolica, Lutzomyia evansi, Lutzomyia flaviscutellata, Lutzomyia gomezi, Lutzomyia hartmanni, Lutzomyia intermedia, Lutzomyia llanosmartinsi, Lutzomyia longipalpis, Lutzomyia migonei, Lutzomyia panamensis, Lutzomyia paraensis, Lutzomyia peruensis, Lutzomyia pessoai, Lutzomyia trapidoi, Lutzomyia umbratilis, Lutzomyia wellcomei, Lutzomyia whitmani, Lutzomyia ylephiletor, Lutzomyia yucumensis, Lutzomyia olmeca, Phlebotomus alexandri, Phlebotomus ansarii, Phlebotomus argentipes, Phlebotomus ariasi, Phlebotomus caucasicus, Phlebotomus celiae, Phlebotomus chinensis, Phlebotomus duboscqi, Phlebotomus kandelakii, Phlebotomus langeroni, Phlebotomus longicuspis, Phlebotomus longiductus, Phlebotomus longipes, Phlebotomus martini, Phlebotomus near rossi, Phlebotomus neglectus, Phlebotomus orientalis, Phlebotomuspapatasi, Phlebotomus pedifer, Phlebotomus perfiliewi, Phlebotomus perniciosus, Phlebotomus salehi, Phlebotomus sergenti, Phlebotomus smirnovi, Phlebotomus tobbi, Phlebotomus transcaucasicus, or Phlebotomus vansomeranae.

31. The method of claim 5, wherein the flea is Aetheca wagneri, Amphipsylla sp., Anomiopsyllus sp., Atyphloceras sp., Callopsylla sp., Catallagia sp., Ceratophyllus sp., Chiastopsylla sp., Citellophilus sp., Coptopsylla sp., Craneopsylla sp., Ctenocephalides sp., Ctenophthalmus sp., Diamanus montanus, Dinopsyllus sp., Eumolpianus eumolpi, Foxella sp., Frontopsylla sp., Hectopsylla sp., Hoplopsyllus sp., Hystrichopsylla sp., Listropsylla sp., Malaraeus sp., Megabothris sp., Megarthroglossus sp., Meringis sp., Monopsyllus sp., Neopsylla sp., Neotyphloceras sp., Nosopsyllus sp., Odontopsyllus sp., Opisocrostis sp., Opisodasys sp., Orchopeas sp., Oropsylla sp., Parapsyllus sp., Pleochaetis sp., Polygenis sp., Psocopsylla sp., Pulex irritans, Rhadinopsylla sp., Stivalius sp., Synosternus sp., Thrassis sp., Tiamastus sp., Tritopsylla sp., Xenopsylla cheopis, or Xenopsylla sp.

32. The method of claim 5, wherein the louse is a member of a Haematopinidae, Hoplopleuridae, Linognathidae, Pediculidae, Polyplacidae, or Pthiridae family.

33. The method of claim 32, wherein the member of the Haematopinidae, Hoplopleuridae, Linognathidae, Pediculidae, Polyplacidae, or Pthiridae family is Haematopinus asini, Haematopinus eurysternus, Haematopinus quadripertusus, Haematopinus suis, Haematopinus tuberculatus, Haematopinus tuberculatus, Haemodipsus ventricosus, Hoplopleura captiosa, Hoplopleura pacifica, Linognathus africanus, Linognathus africanus, Linognathus ovillus, Linognathus pedalis, Linognathus setosus, Linognathus stenopsis, Linognathus vituli, Pediculus humanus capitis, Pediculus humanus humanus, Polyplax serrata, Polyplax spinulosa, Pthirus pubis, or Solenopotes capillatus.

34. The method of claim 5, wherein the triatomine bug is Panstrongylus megistus, Rhodnius prolixus, Triatoma brasiliensis, Triatoma dimidiate, or Triatoma infestans.

35. The method of claim 1, wherein the PCR primers specific for at least three blood meal hosts species hybridize with a satellite DNA, a tandem repeat, or a mitochondrial gene from the blood meal host species.

36. The method of claim 1, wherein the arthropod blood-meal host species is an ape, a bat, a bear, a bird, a bovine, a camelid, a canid, a cervid, a dog, an equid, a felid, a fox, a giraffe, a goat, a horse, a human, a monkey, an opossum, a pig, a rabbit, a raccoon, a rodent, a sheep, a shrew, or a wild boar.

37. The method of claim 36, wherein the bird is a chicken, a crane, a duck, an emu, a goose, a pheasant, a quail, or a turkey.

38. The method of claim 36, wherein the bovine is an auroch, a bison, a buffalo, a domestic cow, an eland, a four-horned antelope, a guar, a spiral-horned antelope, a waterbuck, or a yak.

39. The method of claim 36, wherein the camelid is an alpaca, a Bactrian camel, a dromedary, a guanaco, a llama, or a vicuna.

40. The method of claim 36, wherein the canid is a coyote, a dog, a dog-wolf hybrid, a jackal, or a wolf.

41. The method of claim 36, wherein the cervid is a brocket, a chital, an elk, a montjac, a moose, a mule deer, a red deer, a reindeer, a roe deer, a seka deer, or a white-tailed deer.

42. The method of claim 36, wherein the equid is a donkey, a horse, a mule, or a zebra.

43. The method of claim 36, wherein the felid is a bobcat, a cougar, a domestic cat, a jaguar, a leopard, a lion, a lynx, a mountain lion, an ocelot, a panther, a puma, or a tiger.

44. The method of claim 36, wherein the rodent is a capybara, a chipmunk, a gerbil, a marmot, a mouse, a prairie dog, a rat, a squirrel, or a vole.

45. The method of claim 44, wherein the mouse is a North American deer mouse or a whitefooted mouse.

46. The method of claim 1, wherein the extended products are analyzed by mass spectrometry.

47. The method of claim 46, wherein the mass spectrometry is MALDI-TOF mass spectrometry.

48. The method of claim 1, wherein the PCR primers are selected from nucleotide sequences set forth in SEQ ID Nos: 2, 3, 10, 11, 12, 13, 15, 16, 20, 21, 23, 24, 25, 26, 28, 29, 33, 34, 40, 41, 42, 44, 46, 47, 53, 54, 57, 60, 61, 62, 63, 69, 70, 74, 75, 81, 82, 84, 85, 86, 89, 90 91, 92, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 107, 108, 109, 111, 112, 115, 116, 118, 119, 120, 122, 123, 126, 127, 128, 129, 131, 132, 134, 136, 137, 138, 139, 140, 141, 142, 144, 145, 146, 148, 149, 151, 152, 153, 154, 156, 157, 160, 161, 166, 167, 169, 170, 173, 174, 178, 179, 180, 181, 182, 183, 186, 187, 188, 189, 194, 195, 200, 201, 203, 204, 206, 207, 209, 210, 212, 213, 215, 216, 218, 219, 221, 222, 224, 225, 226, 228, 229, 231, 232, 236, 237, 239, 240, 242, 243, 244, 245, 248, 249, 252, 253, 256, 257, 260, 261, 263, 264, 265, 266, 269, 270, 273, 274, 277, 278, 280, 281, 282, 283, 286, 287, 289, 290, 293, 294, 298, 299, 302, 303, 305, 306, 307, 308, 310, 311, 315, 316, 320, 321, 324, 325, 328, 329, 332, 333, 336, 337, 339, 340, 341, 342, 346, 347, 348, 349, 351, 352, 355, 356, 357, 358, 359, 360, 361, 362, 366, 367, 368, 369, 373, 374, 375, 376, 378, 379, 381, 382, 385, 386, 388, 389, 390, 392, 393, 397, 398, 401, 402, 403, 404, 407, 408, 411, 412, 415, 416, 417, 418, 421, 422, 425, 426, 427, 428, 429, 430, 433, 434, 436, 437, 438, 439, 443, 444, 445, 446, 450, 451, 452, 453, 454, 456, 457, 458, 462, 463, 464, 465, 468, 469, 472, 473, 476, 477, 480, 481, 484, 485, 487, 488, 490, 491, 494, 495, 498, 499, 501, 502, 504, 505, 507, 508, 509, 510, 511, 512, 514, 515, 518, 519, 520, 522, 523, 524, 525, 529, 530, 531, 532535, 536, 538, 539, 541, 542, 543, 544, 546, 547, 548, 549, 553, 554, 555, 556, 558, 559, 560, 561, 562, 563, 566, 567, 568, or 569.

49. The method of claim 1, wherein the extension primers are selected from nucleotide sequences set forth in SEQ ID NOs:1, 8, 9, 14, 19, 22, 30, 31, 32, 39, 43, 45, 48, 49, 50, 52, 55, 58, 64, 65, 66, 67, 68, 71, 72, 76, 80, 83, 88, 93, 106, 110, 113, 114, 117, 121, 124, 125, 130, 133, 135, 143, 147, 150, 155, 158, 159, 165, 168, 172, 175, 177, 184, 185, 192, 193, 199, 202, 205, 208, 211, 214, 217, 220, 223, 227, 230, 235, 238, 241, 246, 247, 250, 251, 254, 259, 262, 268, 272, 276, 279, 285, 288, 292, 296, 297, 301, 304, 309, 314, 317, 319, 322, 323, 330, 331, 335, 338, 343, 344, 345, 350, 353, 354, 363, 364, 365, 370, 371, 372, 377, 380, 383, 391, 394, 396, 399, 405, 409, 413, 419, 423, 424, 431, 432, 435, 440, 441, 442, 447, 449, 455, 459, 461, 466, 471, 475, 479, 483, 486, 489, 493, 496, 497, 500, 503, 506, 513, 517, 521, 526, 527, 534, 537, 540, 545, 550, 552, 557, 564, or 565.

50. The method of claim 1, wherein the amplified products or the extended products are analyzed by next generation sequencing.

51. The method of claim 50, wherein the amplified products are selected from nucleotide sequences set forth in SEQ ID Nos: 4, 5, 6, 7, 17, 18, 27, 35, 36, 37, 38, 51, 56, 59, 73, 77, 78, 79, 87, 104, 105, 162, 163, 164, 171, 176, 190, 191, 196, 197, 198, 233, 234, 255, 258, 267, 271, 275, 284, 291, 295, 300, 312, 313, 318, 326, 327, 334, 384, 387, 395, 400, 406, 410, 414, 420, 448, 460, 467, 470, 474, 478, 482, 492, 516, 528, 533, or 551.

52. The method of claim 1, wherein the biological sample is a biopsy, a blood sample, a cerebral spinal fluid (CSF) sample, an environmental sample, a fluid from a skin blister or ooze from a lesion, a milk sample, a saliva sample, a sewage sample, a synovial fluid sample, a tick homogenate, a tissue sample, or a urine sample.

53. The method of claim 52, wherein the blood sample is a plasma sample, a peripheral blood mononuclear cell (PBMC) sample, a serum sample, or a whole blood sample.

54. The method of claim 53, wherein the method is able to identify an arthropod-borne pathogen in a 1 mL blood sample if greater than 10 arthropod-borne pathogen genomes are present in the blood sample.

55. The method of claim 1, wherein multiple arthropod-borne pathogens are present in the biological sample.

56. The method of claim 1, wherein the biological sample is a pooled sample.

57. The method of claim 1, wherein the biological sample is a sample from a single subject.

58. The method of claim 1, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 24 hours.

59. The method of claim 58, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 12 hours.

60. The method of claim 58, wherein the method is able to provide identification of the arthropod species, the arthropod-borne pathogen, and the blood meal host in less than 8 hours.

61. A method for diagnosing an arthropod-related disease which comprises:(a) identifying the arthropod and the arthropod-borne pathogen in the biological sample from a human patient or an animal subject by the method of claim 1; and (b) using the identification of the arthropod and the arthropod-borne pathogen to diagnose the arthropod-related disease.

62. The method of claim 61, wherein the arthropod-related disease is African swine fever, Alkhurma hemorrhagic fever, anaplasmosis, babesiosis, bluetongue disease, chikungunya, dengue, ehrlichiosis, encephalitis, equine encephalitis, hepatozoonosis,Lyme disease, malaria, Oropouche fever, plague, rickettsiosis, Rocky Mountain Spotted Fever, tick-borne relapsing fever, tularemia, West Nile, yellow fever, or zika fever.

63. The method of claim 61, wherein the human patient is a patient who (i) experienced an arthropod bite; (ii) is suspected of having been bitten by an arthropod; or (iii) is experiencing fever / chills, joint pain, muscle aches, persistent fever, or rash.

64. The method of claim 61, wherein the animal subject (i) experienced an arthropod bite;(ii) is suspected of having been bitten by an arthropod; or (iii) is showing signs of lethargy, lameness, fever, joint pain or swelling, or swollen lymph nodes.

65. The method of claim 61, wherein the arthropod-borne disease is an acute or a chronic arthropod-borne disease.

66. A method of disease vector surveillance which comprises analyzing biological samples by the method of claim 1 in order to identify an arthropod species or an animal host that may act as disease vectors.

67. A method of sentinel surveillance which comprises analyzing biological samples from a sentinel source by the method of claim 1 in order to identify the arthropod species, the arthropod-borne pathogen(s), and / or the blood meal hosts in the sentinel source.

68. A method of monitoring an eradication program which comprises analyzing biological samples by the method of claim 1 in order to identify the arthropod species, the arthropod-borne pathogen(s), and the blood meal hosts.

69. A kit for identification of an arthropod species, an arthropod-borne pathogens, or a blood meal hosts from a single biological sample, wherein the kit comprises:(a) a set of polymerase chain reaction (PCR) primers to generate amplified products, wherein the set of PCR primers comprise (i) PCR primers specific for at least ten arthropod-borne pathogens, (ii) PCR primers specific for at least three species of arthropods, and (iii) PCR primers specific for at least three host species;(b) a set of extension primers; and(c) appropriate reagents and instructions for a user to identify the arthropod-borne pathogen(s), the arthropod species, and the blood meal host(s).

Images