Devices and methods for screening a disease vector

Abstract

Self-enclosed devices for mechanical processing of disease vectors and related screening techniques are disclosed. The devices and methods including lateral flow assays may be particularly useful in at-home screening for Lyme disease.

Description

[0001]Docket No. L2117-7001WO DEVICES AND METHODS FOR SCREENING A DISEASE VECTOR CROSS REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Patent Application No.63 / 687,561, filed ^^ August 27, 2024 and U.S. Patent Application No.63 / 689,267, filed August 30, 2024, the entire disclosure of each of which is hereby incorporated herein by reference in its entirety for all purposes. FIELD OF TECHNOLOGY ^^^ One or more aspects relate to devices and methods for screening a disease vector. BACKGROUND Tick and other pest-borne diseases are a serious and growing problem, with a positive diagnosis of Lyme disease now recorded in every state of the United States of America. A ^^^ shorter time to diagnosis for these diseases correlates to higher rates of successful treatment of the diseases. Current diagnostic tests for Lyme disease have a delay of several weeks to diagnosis, as it takes time for a human immune system to produce enough antibodies to be sensed by current diagnostic tests. ^^^ SUMMARY In accordance with one or more aspects, a device for processing a disease vector for screening is disclosed. The device may include a receptacle defining a crushing recess constructed and arranged to receive a disease vector for processing, a cap configured to create a self-contained system and not reopen when securably mated with the receptacle while still ^^^ being rotatable relative to the receptacle to facilitate processing of the disease vector within the self-contained system, and a crushing element extending from the cap into the receptacle and configured to engage with the crushing recess via rotation of the cap to process the disease vector. The crushing element may further comprise a cam-guided plunger system with ^^^ unidirectional locking, permitting initial rotational engagement without downward force. Annular rib grinding surfaces with saw-tooth profiles may be incorporated on the crushing recess or crushing head to enhance homogenization. One or more openings may be provided between the grinding chamber and an associated sample or reagent chamber, enabling post- ^1- ^ Docket No. L2117-7001WO crushing mixing of extracted material with reagent fluid without breaching the self-contained system. In some aspects, processing the disease vector may involve at least partially fragmenting the disease vector. The self-contained system may be configured to exert a ^^ mechanical force on the disease vector. The self-contained system may be configured to exert mechanical force on the disease vector in two axis. The crushing head may be guided by cam followers engaged with a barrel cam track within the receptacle wall, enabling progressive engagement of the grinding surfaces. The grinding surfaces may include annular ribs with saw-cut edges oriented to maximize shear ^^^ forces and ensure full rupture of the exoskeleton. The cap may incorporate a one-way snap-fit lock preventing reopening after closure, and a dedicated port configured for insertion of a lateral flow or other diagnostic test strip, thereby permitting in-situ analysis without contaminant release. Ergonomic design features, such as contoured gripping surfaces and anti-slip textures, may be included to facilitate safe, one-handed field operation by users with ^^^ minimal training. In some aspects, at least one of the crushing recess and the crushing element define a surface roughness to facilitate processing of the disease vector. The crushing element may comprise a crushing head connected to a crushing implement. The crushing element may comprise one or more contact portions to facilitate processing of the disease vector. ^^^ In certain embodiments, one or more openings are defined between the crushing recess and a reagent chamber, allowing controlled fluid transfer following crushing. This configuration enables homogenized biological material to be automatically mixed with reagent fluid, eliminating the need for transfer to a secondary vessel. The crushing apparatus may further include ergonomic design features such as balanced weight distribution, finger- ^^^ contoured grips, and textured surfaces to allow reliable operation under field conditions, including use with gloves or in wet environments. In some aspects, the receptacle is further configured to receive fluid to facilitate screening of the processed disease vector. The device may further comprise a fluid chamber. The fluid chamber may be contained within the lid or receptacle. ^^^ In some aspects, the device may further comprise a separation mechanism to facilitate downstream disease vector screening. In some aspects, the device is connectable to an assay device. In at least some aspects, the assay device does not involve a DNA-based assay. The device may be configured to receive an assay test strip to facilitate disease vector screening. ^2- ^ Docket No. L2117-7001WO In some aspects, screening the disease vector may involve a fluoroimmunoassay. In other aspects, the assay device may be a colorimetric assay. Screening the disease vector may involve nanoparticle (NP)-labeled antibody and / or antigen-based sensing. In some aspects, wherein the assay device uses both an anti-B. burgdorferi polyclonal antibody and an anti- ^^ OspA antibody. In some specific non-limiting aspects, the concentration of signal antibody may be about 40 ug / mL. In some aspects, the device is a tick screening device intended for at-home use. In accordance with one or more aspects, a lateral flow assay (LFA) for screening a disease vector is disclosed. The LFA may comprise an anti-B. burgdorferi polyclonal ^^^ antibody and an anti-OspA antibody. In accordance with one or more aspects, a kit is disclosed. The kit may comprise a device for processing a disease vector as described herein. In some aspects, the kit may further comprise a source of a buffer solution. The kit may further comprise a source of labeled antibodies. The source of labeled antibodies may ^^^ comprise a sealed dropper of a buffer solution containing the labeled antibodies. In some aspects, the kit may further comprise an assay device. The assay device may be an antibody and / or antigen-based sensing assay. The assay device may be a fluoroimmunoassay. The assay device may be a colorimetric assay device. In some aspects, the assay device may be a lateral flow assay (LFA). The assay device may involve an LFA ^^^ test strip. In some aspects, the assay device may use both an anti-B. burgdorferi polyclonal antibody and an anti-OspA antibody. The assay device may use an anti-B. burgdorferi polyclonal antibody, an anti-OspA antibody, and an anti-OspC antibody. In non-limiting aspects, a concentration of signal antibody-AuNP present in the LFA sample buffer may be ^^^ about 4 ug / mL. The signal antibody’s antibody to AuNP conjugation ratio may be about 40 ug / mL per 10 OD. In some aspects, the assay device may screen for multiple vector-borne diseases in a single sample using a unique capture-signal antibody sandwich combination for each bacteria tested. ^^^ In some aspects, an LFA test strip may be defined by an average pore size of at least about 6-12 uM. In some non-limiting aspects, the LFA test strip does not include a conjugate pad on which the signal antibody is immobilized. The signal antibody may be present in a buffer solution. ^3- ^ Docket No. L2117-7001WO In some aspects, the assay may screen for a protein associated with Anaplasma phagocytophillium or Babesia microti. In some aspects, the assay may be an aggregation-based assay. In some aspects, the kit may further comprise a base configured to stabilize the ^^ receptacle during processing of the disease vector. In some aspects, the kit may be intended for at-home use. In some aspects, the kit may specifically be a Lyme disease screening kit. In accordance with one or more aspects, a method of screening a disease vector is disclosed. The method may involve steps of processing the disease vector using a device as ^^^ described herein to produce a solution, and subjecting the solution to an assay to assess the presence or absence of a disease-causing pathogen or molecule causing an immune response. In some aspects, processing the disease vector may involve applying mechanical force across two axis. In some aspects, the assay may not be a DNA-based assay. ^^^ In some aspects, the assay may screen for a target protein associated with the disease- causing pathogen or molecule causing an immune response. In non-limiting aspects, the disease-causing pathogen or molecule causing an immune response may be associated with Lyme disease, babesiosis, ehrlichiosis, Rocky Mountain Spotted Fever, anaplasmosis, Southern Tick-Associated Rash Illness, Tick-Borne Relapsing Fever, and tularemia. ^^^ In some aspects, the target protein may be associated with a Lyme disease-causing bacteria. The target protein may be associated with B. burgdorferi. The assay may target a B. burgdorferi protein. In some non-limiting aspects, the assay may target outer surface protein A (OspA). In other aspects, the assay may target a tick salivary gland protein. In some non-limiting aspects, the target protein may be associated with a non-Lyme ^^^ disease tick-borne disease-causing pathogen. In some non-limiting aspects, the assay may target Babesia microti surface antigen 1 (BmSA1) protein, Babesia microti glycosylphosphatidylinositol-anchored protein 12 (BmGPI12) protein, and Anaplasma phagocytophilum major surface protein-2 (Msp2) protein. In some non-limiting aspects, the assay may target a non-protein molecule. In some ^^^ non-limiting aspects, the assay may target galactose-alpha-1,3-galactose (alpha-gal). In some aspects, the assay may be an antibody and / or antigen-based sensing assay. The assay may be a lateral flow assay (LFA). In other embodiments, the assay may be an aggregation-based assay. ^4- ^ Docket No. L2117-7001WO In some aspects, the assay may further screen for a second target protein associated with a second disease-causing bacteria. In accordance with one or more aspects, a lateral flow assay (LFA) for screening a disease vector may be any LFA as described herein. ^^ In some non-limiting aspects, the LFA may screen for a target protein associated with a Lyme disease-causing bacteria. The crushing apparatus may also contain a heating component or element, to allow heating of the contents of the crushed disease vector and / or fluid. The heating element may be powered by a battery internal or external to the crushing apparatus. The heating ^^^ component may be external to the crushing apparatus and be composed of a sleeve or containment that allows for heating of the crushing apparatus contents using a battery or chemical means. The present technology has been developed in response to the current state of the art and, in particular, in response to problems and needs that have not been fully or completely ^^^ solved by currently available disease vector crushing devices or screening methods. Accordingly, it is an object of the present invention to provide a disease vector crusher that can adequately or fully fragment a disease vector, that may include but is not limited to a tick, multiple ticks or small pests, to enable a further test reaction to be effectively conducted on the released components or contents of the disease vector. ^^^ It is an object of the present invention to prove an apparatus for crushing disease vectors that can efficiently crush a disease vector in significantly less time, such as 50% less time or more, than is required by prior art crushing apparatus or methods. It is also an object of the present invention to provide an apparatus for crushing disease vectors that require a lower knowledge barrier to entry as to the technical knowledge ^^^ required by the user, such as the operation of microblenders, “bead beater” vials or centrifuges, that is currently required by prior art crushing apparatus or methods. Additionally, it is an object of the present invention to provide an apparatus for crushing disease vectors in a secure manner and which requires significantly less effort than prior art apparatus, such as with a secured lid and rotating action, so that there is significantly ^^^ lowered risk to the user through the inappropriate use of the prior art apparatus or methods. Further, it is an object of the present invention to provide an apparatus for crushing disease vectors which prevents user contact with the disease vector or one or more pathogens or molecules causing an immune response^carried by the disease vector once the disease ^5- ^ Docket No. L2117-7001WO vector has been placed into the apparatus, such as with a lid that closes a receptacle and does not reopen to form a self-enclosed system. Finally, it is an object of the present invention to provide an apparatus for crushing disease vectors which is quiet and portable such that the disease vector can be crushed in situ, ^^ without requiring access to specific equipment or locations containing such equipment as with prior art apparatus or methods. A feature of the invention which enables the above identified objects to be achieved is a crushing or grinding element containing a crushing head connected to a crushing implement and a crushing recess, between which the disease vector will be placed for crushing. In the ^^^ preferred embodiment, the disease vector placed in the crushing recess will be sandwiched between the crushing recess and the crushing head. The crushing head is rotatably contacted to the crushing recess and is configured such that the contact portions between the crushing head and crushing recess will cause the disease vector to be adequately or fully fragmented with a mechanical force applied, so that the contents of the disease vector can be extracted ^^^ and released. The contact portions between the crushing head and crushing recess may also comprise of surface designs to assist in the crushing or grinding of the disease vector with a mechanical force applied on the outside of the lid and / or receptacle. An exemplary feature of the invention that enables the above identified objects to be achieved is a crushing surface on the crushing recess, the crushing head, or both, configured ^^^ with a rough surface or surface with patterns of alternating recessed and raised portions in between which the disease vector will be sandwiched and a mechanical force is applied for crushing. The receptacle that houses the crushing recess at its base may be a single integral component or several separate components. The disease vector will be placed into the ^^^ crushing recess in the receptacle, and the receptacle shall be designed such that it is difficult for the disease vector to escape once placed into the receptacle. The receptacle will fit to a lid with a slot that is designed to create a self-enclosed system, where the lid cannot reopen once it has been placed onto the receptacle, so as to prevent the escape of the disease vector, and to minimize the risks of contact between the user ^^^ and the disease vector once the disease vector has been placed into the receptacle. The crushing implement with the crushing head may or may not be connected to the lid. The slot on the lid may be sealed with a membrane, film or suitable covering to ensure that the system is self-enclosed, so as to prevent the aerosolization of the contents of the device, or the escape of any of the device’s contents into the environment during processing. A test strip may then ^6- ^ Docket No. L2117-7001WO be inserted, e.g. via puncturing the covering / film of the slot, at a subsequent point of time to facilitate performing a screening test on the processed disease vector. The self-enclosed system will also be secure such that any mechanical force like shaking, pressing, pushing, crushing or grinding, applied to the inside or outside of the receptacle and lid, will not cause ^^ any contents, such as fluid or fragmented disease vector parts or any combination of both, to leak out. There may be fluid that is contained or introduced within the lid, receptacle or any other component of the crushing apparatus to allow for it to contact with the extracted components of the crushed disease vector such that a test reaction can be conducted to test for ^^^ the presence or absence of one or more pathogens or molecules causing an immune response carried by the disease vector. The test reaction can be conducted via a test strip or test device that can be inserted into the slot on the lid, such that a portion of the test strip can contact with the mixture of the fluid and crushed disease vector (fluid-vector) mix and initiate a test reaction that can indicate the presence or absence of one or more pathogens or molecules ^^^ causing an immune response carried by the disease vector. The crushing apparatus may include a filtration device or membrane or any such separator system, that would allow for separation of parts of the crushed disease vector from the target contents or fluids of the disease vector containing one or more disease-causing pathogens or molecules causing an immune response, for a further test reaction to be ^^^ conducted on the extracted contents of the disease vector. The crushing apparatus may also contain a mechanism, such as a connecting joint, notch, channel, combination of microfluidic channels or chambers, or any such similar design features, to allow the crushed disease vector-fluid solution to be accessed by one or more components of a further testing device, such as an assay system, to detect the presence of one ^^^ or more pathogens or molecules causing an immune response. The disease vector crusher can be used by itself for crushing disease vectors, or in conjunction with another testing device to allow for the detection of one or more pathogens or molecules causing an immune response carried by the disease vector. The disease vector crusher can be made of one or any combination of any material (in a non-limiting example, ^^^ resin or plastic). Various methods for processing a disease vector for screening are disclosed. Various assays for screening a processed disease vector are also disclosed. Various test strips associated with assays for screening a processed disease vector are disclosed. ^7- ^ Docket No. L2117-7001WO Various assay techniques for screening a processed disease vector are also disclosed. Various kits for preparing and screening a disease vector are also disclosed. These and other objects and features of the present invention will become more fully apparent from the following description and appended claims, or may be learned by the ^^ practice of the invention as set forth hereinafter. BRIEF DESCRIPTION OF DRAWINGS In order that the manner in which the above-recited and other advantages and objects of the invention are obtained, a more particular description of the invention briefly described ^^^ above will be rendered by reference to a specific embodiment thereof which is illustrated in the appended drawings. Understanding that these drawings depict only a typical embodiment of the invention and are not therefore to be considered to be limiting of its scope, the invention will be described and explained with additional specificity and detail through the use of the accompanying drawings listed hereinbelow. ^^^ FIG.1 is a perspective view of an apparatus of the present invention with the crushing implement and crushing head in a contact or closed position with the crushing recess and the lid enclosed onto the receptacle, forming a self-enclosed system^ FIG.2 is a perspective view of an apparatus of the present invention with an exploded perspective view of the crushing implement and crushing head in an open position, where the ^^^ crushing implement and crushing head have yet to contact with the crushing recess, and the lid has yet to be closed onto the receptacle to form the self-enclosed system^ FIG.3 is a side view of an apparatus of the present invention, which is essentially identical to the apparatus shown in FIG.2, with the apparatus in FIG.3 being in an open position, wherein the crushing implement and crushing head have yet to contact with the ^^^ crushing recess and the lid yet to be enclosed onto the receptacle^ FIG.4 presents a schematic of an apparatus of the present invention with a lateral flow assay strip or test strip inserted, demonstrating the use of the apparatus with the sensing test^ FIG.5 presents various schematic views of the lid of the apparatus of the present invention, from different angles and viewpoints^ ^^^ FIG.6 presents various schematic views of the receptacle of the apparatus of the present invention, from different angles and viewpoints^ FIG.7 presents a schematic of a fluorimmunoassay screening approach in accordance ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^8- ^ Docket No. L2117-7001WO FIG.8 presents a schematic of a mobile application for use in conjunction with the fluorimmunoassay screening approach of FIG.7^ FIG.9 presents a schematic of various labeled antibody and / or antigen-based sensing approaches to screen a disease vector in accordance with one or more embodiments^^ ^^ FIGS.10-13 present schematics of lateral flow assay (LFA) test strips in accordance ^^^^^^^^^^^^^^^^^^^^^^^^^ FIGS.14-15 presents data discussed in accompanying Examples^ FIG.16 presents various schematic views of the exterior of an additional embodiment of the present invention and shows the cam follower and barrel cam track mechanism guiding ^^^ the crushing implement^ FIG.17 is a sectional view illustrating annular ribs with saw-cut profiles and openings for fluid communication between grinding and sample chambers and the communication of these in the two available positions of high (mixing) and low (grinding)^ FIG.18 is a sectional view showing the integration of the 3 components which allow ^^^ for grinding, mixing and test strip insertion^^^^^ FIG.19 is a detail view of the locking cap with integrated test strip port DETAILED DESCRIPTION The present invention is related to an improved apparatus for crushing a disease ^^^ vector, which may include but are not limited to a tick, multiple ticks or one or more other small pest(s) to extract the contents of said disease vector(s). More particularly, the apparatus is a tick or pest crusher which enables a tick, tick(s) or pest(s) to be crushed or its parts separated, to extract specific contents of the tick(s) or pest(s), for a further reaction to be conducted on the extracted contents, such that the presence or absence of one or more ^^^ disease-causing pathogens or molecules causing an immune response can be detected. Current tests for pathogens or molecules causing an immune response in disease vectors like ticks or small pests require a lab test setting, in which the disease vector has to be homogenized for a test reaction to be conducted, to ascertain whether or not pathogens or molecules causing an immune response are carried by the disease vector. This is usually done ^^^ with specialized devices or using technology to homogenize the disease vector. However, there is no readily available mechanical crusher that will allow a user to crush a tick, small pest or disease vector on-the-go, with placement of the disease vector into the mechanical crusher and a mechanical force to be imparted on the disease vector to crush it, through the design of the crusher. ^9- ^ Docket No. L2117-7001WO Crushing of the disease vector, which includes but is not limited to a tick, multiple ticks or small pests, can involve imparting a defect in the exterior of the disease vector, separating some or all parts of the exterior of the disease vector from the internal contents of the tick, and / or fragmentation of the disease vector, to allow exposure of the inner ^^ components of the disease vector. Existing test kits involve the use of a stick with a sharp end in a receptable to enable the user to manually exert a force on the disease vector to impart a defect on its exterior for a further test to be done. However, this manual crushing may not be effective in releasing the contents of the vector, there is a risk of escape of the disease vector if the receptacle is too ^^^ shallow, and with the outcome varying according to user and the size of the disease vector. In the laboratory setting, there are devices such as micro blenders or “bead beater” vials comprising of a small bead or bead(s) in a vial that is shaken vigorously in a device such as a high-powered centrifuge, to pulverize a disease vector. However, such crushing methods requires the operator to have technical knowledge in operating the machinery, the necessary ^^^ components for the pulverization reaction, alongside access to the specialized equipment. In accordance with one or more embodiments, an apparatus for crushing disease vectors that may include but are not limited to a tick, multiple ticks or other small pests, which can adequately or fully fragment a disease vector such that a further test reaction can be conducted on the extracted contents of the disease vector is disclosed. ^^^ In accordance with one or more embodiments, an apparatus for crushing disease vectors that can efficiently crush a disease vector in significantly less time than is required by prior art crushing apparatus or methods as well is disclosed. In accordance with one or more embodiments, an apparatus for crushing disease vectors that require a lower knowledge barrier to entry as to the technical knowledge required ^^^ by the user, that is currently required by prior art crushing apparatus or methods is disclosed. In accordance with one or more embodiments, an apparatus for crushing disease vectors in a secure manner and which requires significantly less effort than prior art apparatus such that there is significantly lowered risk to the user through the inappropriate use of the prior art apparatus or methods is disclosed. ^^^ In accordance with one or more embodiments, an apparatus for crushing disease vectors which prevents user contact with the disease vector or one or more pathogens or molecules causing an immune response carried by the disease vector once the disease vector has been placed into the apparatus is disclosed. ^10- ^ Docket No. L2117-7001WO In accordance with one or more embodiments, an apparatus for crushing disease vectors which is quiet and portable such that the disease vector can be crushed in situ, without requiring access to specific equipment or locations containing such equipment as with prior art apparatus or methods is disclosed. ^^ In accordance with one or more embodiments, an apparatus for the processing of a disease vector may render any associated target bacteria, virus, or parasite (and related surface proteins) or other antigen available for effective screening. The target bacteria,virus, or parasite and related surface proteins or antigens may be associated with one or more biological systems of the disease vector and be made available for screening via adequate ^^^ processing of the disease vector as described herein. There is a need for an improved disease vector crusher that overcomes or avoids the above problems as disclosed herein. There is also a need for assay techniques for effectively and efficiently screening a processed disease vector as disclosed herein. ^^^ There is still further a need to for kits to facilitate the processing and screening of disease vectors as disclosed herein. Devices: The present invention is directed to an improved apparatus for mechanically ^^^ processing disease vectors, including but not limited to a tick, multiple ticks or small pests. More particularly, the apparatus is a disease vector crusher which enables disease vectors to be crushed or its parts separated, to extract specific contents of the disease vector, for a further test reaction to be conducted on the extracted disease vector contents, such that the presence or absence of one or more disease-causing pathogens or molecules causing an ^^^ immune response can be detected. In at least some embodiments, processing of a disease vector may render any associated target bacteria, virus, or parasite (and related surface proteins) or other antigen available for effective screening. The target bacteria, virus, or parasite and related surface proteins or antigens may be associated with one or more biological systems of the disease vector and be made available for screening via adequate ^^^ processing of the disease vector as described herein. In accordance with one or more embodiments, the device may be operable to apply sufficient torque and / or shear force to grind a target vector of any size and species. The device may be effectively operable by any user across a wide spectrum of age, strength and ability. ^11- ^ Docket No. L2117-7001WO In accordance with one or more embodiments, devices achieve effective exoskeleton rupture. In accordance with one or more embodiments, the devices leverage controlled shear force application to ensure sufficient destruction of pathogens. Determination of an optimized ^^ shear force may depend on various factors including but not limited to properties of the disease vector’s exoskeleton, a user’s strength and hand force (e.g. age dependent, and the translation of applied force into shear stress. The shear force needed to rupture a tick’s exoskeleton depends on hydration, tick species, and maturity. ^^^^^^^^^^^^ ^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^!!"^ ^^^^^^^^^ ^^^^^^ #^^^^^^^$^^^^ ^^^^^^^^^^^^^^^^^^^^ ^^ ^^^^^^ %^^ ^^^^^^^^^^^^^^^^^^^^^ ^^%^^^^^^ ^^^%^^^^^^^^ ^^^ !^^^^^"^^^^^^^^ &^^^^^^^^^ ^^^^^^^^^^^^ ^^##^ ^^^ In some embodiments, the grinding surfaces create a high-friction interface to induce shear forces between 50-1000 N, effectively rupturing a disease vector (e.g. tick)’s exoskeleton. The crushing mechanism allows users of varying hand strength to generate sufficient pressure to reach the necessary shear stress of 5-50 MPa. ^^^ In accordance with one or more embodiments, the devices leverage an optimized grinding surface roughness to enhance mechanical engagement for grinding of the target vector and for maintaining durability and efficiency. In some embodiments, the devices may be characterized by a surface roughness of about 30-100 µm Ra. Below is a non-limiting suggested range based on grinding and crushing mechanisms: Surface Type Purpose Suggested Roughness (Ra in µm) Rz (µm) Rt (µm) Concave Grinding Engages tick, applies 100–250 150–400 Surface pressure 20–50 µm µm µm Convex Grinding Crushes tick 150–400 200–600 Surface exoskeleton 30–75 µm µm µm ^^^ • Ra 20–75 µm: Comparable to coarse sandpaper (40-60 grit) or rough-machined surfaces. ^12- ^ Docket No. L2117-7001WO • Rz 100–400 µm: Ensures aggressive material interaction. • Rt 150–600 µm: Allows for effective deformation of the tick exoskeleton. In accordance with one or more embodiments, the devices leverage defined shear ^^ stress thresholds to match biomechanical properties of a target disease vector’s exoskeleton. In some embodiments, a shear stress threshold of about 5-50 MPa may be applicable. In accordance with one or more embodiments, the crushing element may further comprise a cam-guided plunger system with unidirectional locking, permitting initial rotational engagement without downward force. Annular rib grinding surfaces with saw-tooth ^^^ profiles may be incorporated on the crushing recess or crushing head to enhance homogenization. One or more openings may be provided between the grinding chamber and an associated sample or reagent chamber, enabling post-crushing mixing of extracted material with reagent fluid without breaching the self-contained system. In accordance with one or more embodiments, the crushing head may be guided by ^^^ cam followers engaged with a barrel cam track within the receptacle wall, enabling progressive engagement of the grinding surfaces. The grinding surfaces may include annular ribs with saw-cut edges oriented to maximize shear forces and ensure full rupture of the exoskeleton. The cap may incorporate a one-way snap-fit lock preventing reopening after closure, and a dedicated port configured for insertion of a lateral flow or other diagnostic test ^^^ strip, thereby permitting in-situ analysis without contaminant release. Ergonomic design features, such as contoured gripping surfaces and anti-slip textures, may be included to facilitate safe, one-handed field operation by users with minimal training. In accordance with one or more embodiments, one or more openings are defined between the crushing recess and a reagent chamber, allowing controlled fluid transfer ^^^ following crushing. This configuration enables homogenized biological material to be automatically mixed with reagent fluid, eliminating the need for transfer to a secondary vessel. The crushing apparatus may further include ergonomic design features such as balanced weight distribution, finger-contoured grips, and textured surfaces to allow reliable operation under field conditions, including use with gloves or in wet environments. ^^^ The main components of the apparatus shown in FIGS.1–3 include a lid A, receptacle B, crushing implement C, crushing head D, crushing recess E, contact portions F, crushing surface G, slot H. The other design components may include but are not limited to an additional connecting mechanism I, fluid chamber K and filtration mechanism K. The design components I, J and K have not been illustrated in FIGS.1-6. ^13- ^ Docket No. L2117-7001WO FIG.1 shows a perspective view of the apparatus in a contact or closed position, with the crushing head D connected to the crushing implement C, being in contact with the crushing recess E. The disease vector(s) will be placed within the crushing recess E, and will be sandwiched between the crushing head D and the crushing recess E. Additional design ^^ features and mechanisms are situated on the sides of the crushing head D and the crushing recess E are present, with contact portions F, which will assist in the grinding of the disease vector on the crushing surface G, with a mechanical force applied on the exterior of the lid A or the receptacle B. The lid A fits rotatably onto the receptacle B, which will form a self- enclosed system, wherein the lid A can still rotate such that a mechanical force can be applied ^^^ on the exterior of the lid A or receptacle B, to move the crushing head D and crushing recess E rotatably in the interior of the apparatus. The lid A also has the slot H, in which a lateral flow assay or test strip can be inserted to contact with a suspension or fluid-vector mix in receptable B, for a test to be conducted on the suspension or fluid-vector mix. The slot H may be covered by a membrane, film, seal or covering that encloses the entire system and prevents ^^^ the aerosolization of the contents of the device, or the escape of any of the device’s contents into the environment, until the test strip has been inserted into the slot for the test. The self- enclosed system is designed to be secure, such that once the lid A has been fitted onto receptacle B, any mechanical force like shaking, pressing, pushing crushing or grinding applied to the inside or outside of the receptacle and / or lid, will not cause any contents, such ^^^ as fluid or fragmented disease vector parts or any combination of both, to leak out. FIG.2 shows an exploded perspective of the crushing implement C and crushing head D in an open position, relative to the crushing recess E. The crushing implement C is connected to the lid A and the crushing recess E is located in the receptacle B. The crushing implement C and crushing head D have yet to contact with the crushing recess E and the lid ^^^ A has not been closed onto the receptacle B in FIG.2. FIG.3 is a side view of the crushing apparatus in an open position, similar to that of FIG.2. FIG.3 shows that the crushing implement C and crushing head D are connected to the lid A, and can be made of any material (in a non-limiting example, resin or plastic). The various components of the crushing implement C, crushing head D and lid A may or may not ^^^ be made of the same material. FIG.3 also shows that the crushing recess E and crushing surface G is located within the receptacle B. The disease vector shall be placed within the crushing recess E, sandwiched between the crushing surface G and crushing head D. When a downward force is applied onto the lid A to enclose it over the receptacle B, the disease vector shall be caught between the crushing head D and crushing surface G, or within the ^14- ^ Docket No. L2117-7001WO contact portions F. When a mechanical force is applied on the exterior of the lid A or the receptacle B, the disease vector / tick shall be fragmented or crushed more effectively. A suspension or fluid-vector mix is then created as a result of the mechanical force applied on the disease vector / tick, and a test strip inserted through the slot H in the lid A can then enact a ^^ test on the suspension or fluid-vector mix. The various components of the receptacle B, crushing recess E, crushing surface G and contact portions F can be made of any material (in a non-limiting example, resin or plastic), and the various components may or may not be made of the same material. In accordance with one or more non-limiting embodiments, the receptacle B includes ^^^ a convex conical shape in the bottom of the container at an approximate depth at center of 0.1875”-0.5” above the bottom of the container (reference plane). There may be a concave conical face on the crushing head D which matches the geometry of receptacle B to provide a crushing surface when rotated. In an additional embodiment, the crushing surfaces can be level and opposing each other. This crushing surface on the crushing head D may be a ^^^ smooth or roughened surface (0-25µm). Annular saw cuts may be the surface pattern to create the roughened surface. In addition, when rotated, the conical surface is designed to guide the tick / tick particles to teeth at the perimeter of the container conical geometry at a pitch of approximately 0.093”, depth of 0.025” to provide further crushing when rotated. The crushing head D features a reduced number of teeth, primarily intended to catch the tick and ^^^ guide to the exterior teeth on the contact portions F and crushing recess E. The tooth pitch, depth, angle, spacing of the crushing implement C, crushing head D, contact portions F and crushing recess E may be modified to optimize crushing. The objective in processing the disease vector is to enable the contents of the disease vector to be sufficiently extracted and released, such that a further test reaction can be ^^^ conducted on the released components or contents of the disease vector, and the presence or absence of one or more disease-causing pathogens or molecules causing an immune response can be detected. There shall thus be other components that may be worked into the design, which may include but are not limited to an additional connecting mechanism I, fluid chamber J and filtration mechanism K. The design components I, J, K and other potential ^^^ design components are not included in FIGS.1-6, but may be implemented in the crushing apparatus or related kits for the purposes of enhancing the accuracy, reliability and visible reaction of the further test to be conducted on the crushed disease vector components. The connecting mechanism I can be part of the design of the lid A or receptacle B, connected either to the lid A or receptacle B, and allow the crushing apparatus to be connect ^15- ^ Docket No. L2117-7001WO to a further testing system, which may include but is not limited to, an assay system or any other test that will allow for the detection of the presence or absence of one more disease- causing pathogens or molecules causing an immune response in a fluid, fragmented disease vector parts or any combination of both. ^^ The fluid chamber J can be contained within the lid A or receptacle B, or located externally from the crushing apparatus. The fluid chamber J shall contain a fluid that will contact with fragmented disease vector parts to form a suspension or fluid-vector part mix that can be allow for the detection of the presence or absence of one or more disease-causing pathogens or molecules causing an immune response in the suspension or mix, whether ^^^ through a visible reaction in the fluid, suspension, or fluid-vector mix itself, or through the fluid, suspension or fluid-vector mix entering into a further testing system that may include but is not limited to an assay system, that elicits a visible reaction in said further testing system. The filtration mechanism K can be contained within the lid A or receptacle B, or ^^^ located externally from the crushing apparatus. The purpose of the filtration mechanism is to sieve out larger fragmented disease vector components from the fluid, suspension or fluid- vector mix to produce a subsequent fluid, suspension or fluid-vector mix that will provide a more reliable, accurate and visible reaction when it contacts with the assay system. The filtration mechanism will be used after the fluid contacts with the fragmented disease vector ^^^ parts and mixes with it, to produce the subsequent fluid, suspension or fluid-vector mix. These other design components I, J, K and other potential design components can be made of any material (in a non-limiting example, resin or plastic), and the various components may or may not be made of the same material. An exemplary disease vector processing device / crusher / homogenizer and storage ^^^ device comprises but is not limited to a receptacle with an implement or design to allow for ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ and a lid or cover to create a self-enclosed system. The function of the receptacle and the lid is to provide a volume in which a pest, or pests, is held within a self-enclosed system and is not able to escape. The function of the crusher / grinder implement is to impart a defect in the ^^^ exterior of the tick, separate some or all parts of the exterior of the tick from the internal contents of the tick, and / or to crush the tick, to allow exposure of the inner components of the tick. The device will also have a mechanism to either introduce into or contain fluid in the grinding chamber. It will also have a way to allow the crushed tick-fluid solution to be accessed by an assay system to detect the presence of Borrelia burgdorferi. The crusher / ^16- ^ Docket No. L2117-7001WO homogenizer and storage device can be made of any material (in a non-limiting example, resin or plastic). The crusher / homogenizer and storage device can be used by itself, or in conjunction with an assay that screens for the presence of one or more disease-causing pathogen(s) or molecules causing an immune response. ^^ In accordance with one or more embodiments, a lid will enclose the crushing apparatus to form a self-enclosed system in which the disease vector will be stored / contained and which a mechanical force can be applied on the exterior to assist in the crushing of the disease vector. The lid may be comprised of one or more parts. In accordance with one or more embodiments, a receptacle will contain the disease ^^^ vector and form a self-enclosed system with the closure of the lid, such that the disease vector will not be able to escape and a mechanical force can be applied on its exterior to assist in the crushing of the disease vector, which will also allow for a mechanical force to be applied onto the disease vector housed within the receptacle, to crush or fragment it partially or fully. In accordance with one or more embodiments, a crushing implement will be ^^^ connected to the lid and a crushing head, that will allow for the extension of the crushing head to contact with the crushing recess. In accordance with one or more embodiments, a crushing head will be an extension of the crushing implement and be designed in a manner that will rotatably fit and contact with a crushing recess, in which a mechanical force applied on the lid will allow it to crush or ^^^ fragment the disease vector partially or fully. In accordance with one or more embodiments, a crushing recess in which the disease vector will be placed onto, and which will be connected to the receptacle, and will be designed in a manner that will rotatably fit and contact with the crushing head, in which a mechanical force applied onto the receptacle will allow for the crushing or fragmentation of ^^^ the disease vector partially or fully. In accordance with one or more embodiments, contact portions will extend from the crushing head and crushing recess, and be situated at the edges of the crushing head and crushing recess, in which the disease vector can be caught within the contact portions to assist in the grinding of the disease vector on the crushing surface or in the contact portions, when a ^^^ mechanical force is applied on the exterior of the lid or receptacle. In accordance with one or more embodiments, a crushing surface will be located on the crushing recess, wherein the disease vector will be sandwiched between the crushing head and crushing recess when the two components come into contact with each other as the lid is fully enclosed over the receptacle, and which the disease vector will be fully or partially ^17- ^ Docket No. L2117-7001WO fragmented or crushed, when a mechanical force is applied on the exterior of the lid or receptacle. In accordance with one or more embodiments, a connecting mechanism may connect the crushing apparatus and its contents to a further testing system. ^^ In accordance with one or more embodiments, a fluid chamber may contain a fluid that will contact with fragmented disease vector parts to form a fluid, suspension or fluid- vector mix, and which can be contained internally in the crushing apparatus, or introduced externally. In accordance with one or more embodiments, a filtration mechanism may sieve out ^^^ larger fragmented disease vector components from a fluid, suspension or fluid-vector mix to be used after the fluid contacts with fragmented disease vector components. This technology functions as a self-enclosed system in which a tick, or multiple ticks, may be placed and trapped for crushing. The receptacle is designed in such a manner that allows for the tick to be placed ^^^ within it, and have the tick in contact with the crushing surface. To form a self-enclosed system, a lid or cover may be fitted onto the receptacle once the tick is contained in the receptacle. In accordance with one or more embodiments, a unidirectional snap-on lid may be implemented. The unidirectional snap-on lid will allow the lid to be enclosed onto the ^^^ receptacle, but not allow for separation of the two components once the lid has been snapped on. The unidirectional snap-on lid may still be freely rotatable relative to the receptacle once in position to facilitate processing of an enclosed disease vector. The crushing implement may or may not be attached to the lid or other parts of the technology. The crushing implement shall be designed in such a manner within the self- ^^^ enclosed system that it can exert a high enough surface pressure on the tick that a defect can be imparted on the exterior of the tick, and / or to grind / pulverize the exterior of the tick so that the fluids inside can be released, or for the whole tick to simply be homogenized. The technology may include, but is not limited to, being designed in such a manner as to allow for the containment of solution, and may include a filtration device or membrane or ^^^ any such separator system, that would allow for separation of parts of the dissociated or crushed tick from the target fluids or specific contents of the tick containing the target disease-causing pathogens or molecules causing an immune response, for a further assay to be conducted on the extracted contents. A connecting joint, channel, combination of microfluidic channels or chambers, notch or any such similar design features may be present ^18- ^ Docket No. L2117-7001WO in the technology that would connect it to one or more components of the test kit. The technology can be made from any material. This technology comprises but is not limited to a receptacle with an implement or design to allow for the placement of a disease vector, or multiple disease vectors, into the ^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^-enclosed system. The function of the receptacle and the lid is to provide a volume in which a pest, or pests, is held within a self-enclosed system and is not able to escape. The function of the crusher / grinder implement is to impart a defect in the exterior of the tick, separate some or all parts of the exterior of the tick from the internal contents of the tick, and / or to crush the tick, ^^^ to allow exposure of the inner components of the tick. The device will also have a mechanism to either introduce into or contain fluid in the grinding chamber. It will also have a way to allow the crushed tick-fluid solution to be accessed by an assay system to detect the presence of B. burgdorferi. The technology can be made of any material (in a non-limiting example, resin or plastic). This technology could be used by itself, or in conjunction with an assay that ^^^ can screen for the presence of a disease-causing pathogen(s) or molecules causing an immune response. In accordance with one or more embodiments, a crusher for a disease vector, that may include but is not limited to ticks or small pests, which applies pressure to the disease vector to enable a user to quickly and securely crush the tick or small pest(s), without it being able ^^^ to escape once it is placed into the device is disclosed. The disease vector or disease vectors are held within a self-enclosed system with a receptacle secured with a lid configured to snap on. The lid is configured to an extended grinding element containing a crushing head and a crushing recess that will crush the disease vector. The crushing head is rotatably contacted to the crushing recess. Fluid will either be contained or introduced to the disease vector crusher ^^^ and it may contain a connecting channel to connect the fluid to a separate testing device. In accordance with one or more embodiments, devices may generally have structural and / or functional similarities to a salt or pepper grinder, but it may have two axis in which it grinds the disease vector. It will have an “up-down” grinding surface where the tick is crushed and ground vertically by two hard disks. The tick, also as it is squeezed down and ^^^ away from the center, will start being ground by “outer-inner surface” of the outside edge of the rotating disk, and the inside surface of the entire container. Prior to processing, the tick to be crushed typically exhibits a mean particle size (D50) of 5 - 9 mm (nymphs to adults), indicating a relatively coarse structure. After undergoing the improved mechanical treatment, the D50 value was reduced to 100–500 µm (0.1–0.5 mm), ^19- ^ Docket No. L2117-7001WO demonstrating at least a tenfold refinement in particle size. This reduction in mean particle size enhances the surface area-to-volume ratio, leading to improved solubility, better mixing properties, and increased bioavailability. By achieving finer particle sizes with greater crushing efficiency, this method demonstrates a superior approach to mechanical processing ^^ with enhanced performance characteristics. In accordance with one or more embodiments, the grinding surfaces are designed to be rough to promote fragmentation. The size of the grinding surfaces were purposely chosen to be of the correct dimension to ensure break-up of a nymph-sized black-legged tick which is approximately ~1.5 mm or the size of a sesame seed. The grinding surfaces and / or other ^^^ components of the device for processing the disease vector may be tailored to facilitate and optimize a specific use case. In accordance with one or more embodiments, buffer solution may be introduced to the receptacle along with the subject vector before the receptacle is closed and mechanical rotation / grinding force is applied. The buffer solution may generally be used to liberate the ^^^ bacteria from the homogenized tick and then allow it to flow through, e.g. a lateral flow assay strip as discussed herein. In some embodiments, a small slot in the device may have a thin membrane / film or seal which can be removed for the insertion of the LFA. Alternatively, the LFA strip may punch through the slot with membrane / film. ^^^ In accordance with one or more embodiments, the LFA test strip will contact the crushed tick + solution suspension below and the test line will appear above the lid at the portion where the test strip emerges for user visibility. In accordance with one or more embodiments of operation, a tick or other disease vector may be found by a user. The disease vector may be removed from a subject if latched ^^^ and / or engorged. For example, the disease vector may be located in nature, on a clothing garment, on a pet, or on human skin. The tick is then dropped into the open bottom half of the crushing container, and the sample buffer is then poured using a dropper or similar fluid container into the open crushing container. The top lid is then shut onto the bottom lid and “snapped” into place. The two halves of the container are then twisted in opposite directions ^^^ until the tick is satisfactorily processed. Processing may involve imparting defects, dismembering or homogenizing. The processed disease vector may then be subjected to an assay for screening. In some nonlimiting embodiments as described herein, the processed disease vector may be subjected to a lateral flow assay (LFA). The LFA may be punched through the top of the container to access the processed tick / buffer sample, and let sit for a ^20- ^ Docket No. L2117-7001WO period of time, e.g.20 min. Alternatively, the LFA may be inserted through the top of the container through the slot subsequent to a user removing a sticker seal or other protective covering over the slot in order to insert the LFA strip. The LFA may generally stick partially out of the top of the container. After 20 minutes, if there is a visible line on the part of the ^^ LFA that is sticking out the top of the container, the tick that was crushed had B. burgdorferi present in its system. Various components of the device and / or associated kit may depend on or be influenced by the associated assay technique. For example, in embodiments involving LFA, the sample pad on the LFA will act as a sufficient filter rendering an additional filter ^^^ component as described herein unnecessary in the design. Likewise, the processing device may provide sufficient support such that an additional stable base is not required. In accordance with one or more embodiments, a kit may include the following elements: two halves of an open unused tick-crushing container, for example, a receptacle ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^e^^^ squeezed into the tick-^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^ strip. The LFA strip may be in a sterile package. In accordance with one or more alternative embodiments, a disease vector may be placed in a flexible vial. A user may squeeze the vial repeatedly until the tick is sufficiently crushed. The flexible vial may be full of solution buffer (such as PBS) during processing of ^^^ the disease vector. Alternatively, buffer solution may be added subsequent to processing. FIG. 4 generally illustrates this approach. In certain preferred embodiments, the crushing implement is mounted on cam followers that travel along a barrel cam track within the receptacle wall. This arrangement allows the crushing head to rotate freely at the start of engagement without requiring ^^^ downward pressure, then progressively engages the grinding surfaces as the plunger advances. The grinding surfaces may comprise annular ribs with saw-tooth cuts oriented to maximize shear forces and ensure full rupture of the disease vector’s exoskeleton. One or more openings are formed between the grinding chamber and a sample / reagent ^^^ chamber, enabling controlled fluid transfer once crushing is complete. This allows immediate mixing of the extracted biological material with a reagent fluid stored in the device, without opening the enclosure. ^21- ^ Docket No. L2117-7001WO The cap may incorporate a one-way snap-fit lock preventing removal after closure. A dedicated port in the cap is configured for insertion of a lateral flow or other diagnostic test strip, permitting on-site analysis without contaminant release. Ergonomic features may include finger-contoured grips, anti-slip surface textures, ^^ and a geometry that permits comfortable one-handed use by gloved or ungloved operators in outdoor conditions. Alternatively, the tick crusher may include a component that integrates tightly with the lid that allows the buffer solution to be introduced to the receptacle. This shall fit snugly into the lid component and contain a slot that allows for introduction of the test strip to a ^^^ uniform depth into the solution. The mechanism of rotation may include a track that allows for the lid component to provide vertical movement providing for mixing of the buffer solution in addition to the grinding component. ^^^ Screening Techniques: To shorten time to treatment, instead of waiting for bitten humans to produce antibodies in response to an infection, one could test the disease vector, such as but not limited to ticks, biting flies, and mosquitoes, that delivered the bite for the presence of disease-causing pathogens or microbial agents, such as but not limited to bacteria, viruses, ^^^ parasites or molecules causing an immune response. While any disease vector can be screened for any pathogen or microbial agent or molecules causing an immune response that can be carried by said disease vector in accordance with various embodiments disclosed herein, one non-limiting example which may be used throughout merely for descriptive simplicity is ticks carrying the Lyme disease-causing bacteria B. burgdorferi. ^^^ In accordance with one or more embodiments, a disease vector can be screened for a tick-borne disease such as but not limited to Lyme disease, Ehrlichiosis, Babesiosis and Anaplasmosis. In accordance with one or more embodiments, a disease vector may be screened for one or more diseases or disease-causing antigens selected from the group consisting of: ^^^ Borrelia sp., Ehrlichia sp., Anaplasma sp., Rocky Mountain spotted fever, Rickettsia sp., tularemia, Babesia sp., heartland virus, bourbon virus, Colorado tick fever, Powassan virus disease, relapsing fever, Mediterranean spotted fever, R. conorii, tick-borne encephalitis (TBE Virus), neoehrlichiosis, Lyme-like illness, and molecular-based antigens such as galactose-alpha-1,3-galactose causing tick-borne alpha-gal syndrome. ^22- ^ Docket No. L2117-7001WO In accordance with one or more embodiments, a disease vector may be screened for a panel of diseases, such as a group of diseases associated with a certain geographic region. In accordance with one or more embodiments, a screening device such as but not limited to a lateral flow assay (LFA) may be operable to effectively detect one or more of the ^^ pathogens or microbial agents disclosed herein as well as one or more other diseases which may be associated with a subject disease vector. In some embodiments, the screening device may provide for a screening panel, such as a regional screening panel. In accordance with one or more embodiments, devices, kits and methods can be used to test a disease vector such as ticks for the presence of a disease such as Lyme disease. ^^^ In accordance with one or more embodiments, a screening device may perform in the field of Lyme disease detection. Currently, if a user finds an engorged tick on their body or on a pet or other person, they have limited ways to determine if the tick they were bitten with is carrying the Lyme disease-causing bacteria, B. burgdorferi. The current main method of screening a tick for Lyme disease is mailing the tick to a lab and waiting for the lab to mail ^^^ back a tick DNA result. There is currently no device available to the general public that can screen a discovered tick for the presence of B. burgdorferi without mailing the sample to a lab to run DNA testing. In accordance with one or more embodiments, a discovered tick can be tested for the presence of the target pathogen or microbial agent or molecules causing an immune response without mailing samples to a lab. ^^^ Conventional tests for diagnosis of Lyme disease included polymerase chain reaction (PCR) which is an expensive and time-consuming process. In accordance with one or more embodiments, a screening technique which is not a DNA-based assay as done in labs for similar testing may be implemented. Various proteins which support persistence of B. burgdorferi, resulting in Lyme ^^^ disease, are known to those skilled in the art. Such proteins include B. burgdorferi proteins or Tick salivary gland proteins: B. burgdorferi proteins: Lipoproteins, Outer surface proteins (Osps), BB0323, BBA52, and BBA64, Dps, VlsE, lipoproteins, Lp6.6, LA7, BptA, BBA07, bbe31, BbCRASP proteins, ^^^ BB0323, BBA52, BBA64, BmtA (BB0219), Enzymes in B. burgdorferi, BB0646 Tick salivary gland protein: Salp15, Salp25D, Salp20, Tick histamine release factor, Tick Salivary Lectin Pathway Inhibitor (P8), TRE31, TROSPA (tick receptor for OspA) ^23- ^ Docket No. L2117-7001WO In accordance with various embodiments, screening approaches may use antibodies which target one or more of these unique proteins. For example, labelled antibodies may target one or several antigens from among the above list of unique proteins. If the antigen(s) are present, the antibodies will bind, and it will result in a color change of the solution. This ^^ color change will indicate to the user that the protein, and by extension B. burgdorferi, is present. There is no intention to limit the ensuing ideas to only within the scope of a disease vector test kit. In a non-limiting example, for example, they may be used in an at-home test kit for a bacteria on human skin. ^^^ Furthermore, it is envisioned that the screening results can contribute to identify hot spots where the Lyme disease is particularly present. Location details can be gathered, data can be anonymized, analyzed and shared with health departments to maximize awareness and help allocating resources in relevant areas. The only information shared may be location and positive or negative results, obviating any need for HIPAA compliance. ^^^ Potential extensions to cover other pathogens (e.g. viruses) or molecules causing an immune response spread by disease vectors such as mosquitoes (e.g. Zika) is also envisioned. Fluoroimmunoassay technology: In accordance with one or more embodiments, a fluoroimmunoassay approach may be ^^^ used for screening a disease vector. When a tick is found on a person or pet, it may be processed in accordance with the devices and methods described herein. Buffer may be added prior or subsequent to mechanical processing. A user may then pour a second vial of solution containing a buffer (such as PBS) and labeled antibodies into the first vial containing the crushed up tick. After a ^^^ short period of time, the solution will change color if the targeted unique protein supporting persistence of B. burgdorferi is present, indicating the tick was a carrier of Lyme disease. FIG.7 generally references this approach. A related kit may include a device for processing the subject disease vector as described herein, a source of solution buffer to facilitate vector processing, and a sealed ^^^ dropper filled with buffer and specific labeled antibodies. A stable base may optionally hold the tick vial during screening. In accordance with one or more embodiments, a disease vector may be screened for one or more diseases or disease-causing antigens selected from the group consisting of: Borrelia sp., Ehrlichia sp., Anaplasma sp., Rocky Mountain spotted fever, Rickettsia sp., ^24- ^ Docket No. L2117-7001WO tularemia, Babesia sp., heartland virus, bourbon virus, Colorado tick fever, Powassan virus disease, relapsing fever, Mediterranean spotted fever, R. conorii, tick-borne encephalitis (TBE Virus), neoehrlichiosis, Lyme-like illness, and molecular-based antigens such as galactose-alpha-1,3-galactose causing tick-borne alpha-gal syndrome. ^^ In accordance with one or more embodiments, a companion mobile-based application may use a mobile phone to identify a subtle color change in a given solution as illustrated in FIG.8. The object containing the tick and antibody solution will be placed inside a provided container to block extraneous environmental light. The mobile phone with a flashlight and a camera will be placed on top of the container. There will be a hole for the camera and ^^^ flashlight on the phone to illuminate the contents of the container. A magnet may optionally facilitate proper placement of the mobile phone. The phone will take an initial scan of the solution, then an additional scan of the solution at a second period of time (e.g. five minutes later). The mobile app will compare the images for a color change in the solution. It will then report to the user if a color change occurred within the solution. If so, the user will know that ^^^ Lyme disease-causing bacteria were present in their tick sample. Colorimetric assays: In accordance with one or more embodiments, proteins which support persistence of B. burgdorferi in tick samples may be detected by colorimetric nanoparticles. ^^^ In accordance with one or more embodiments, Nanoparticle (NP)-labeled antibodies and / or antigen-based sensing may be implemented. In accordance with one or more embodiments, nanoparticles (and / or nanorods) may be utilized to detect different kinds of proteins. Certain proteins support persistence of B. burgdorferi, resulting in Lyme disease. The proteins could come from either B. burgdorferi ^^^ proteins or Tick salivary gland proteins as discussed and identified above. This screening approach is generally illustrated in FIG.9. In accordance with one or more embodiments, a molecule with high affinity with the representative protein may be conjugated to nanoparticles or nanorods with intrinsic colorimetric properties. Upon binding, the emission color changes due to the alteration in the^^^ medium. Beneficially, the color change can be seen by naked eyes of a user through a take- home kit, resulting in a quick approach for predicting the likelihood of Lyme disease. In accordance with one or more embodiments, this technology consists of a detection agent that includes but is not limited to nanoparticle (NP)-labeled antibodies and / or antigens that will be able to sense the presence of B. burgdorferi in ticks. Antibodies and antigens ^25- ^ Docket No. L2117-7001WO could be used in a variety of systems to sense for targets including but not limited to an ELISA-like lateral flow assay or an in-solution aggregation-based color change. It also could be used to sense for the presence of multiple bacteria and viruses at once. ^^ Nanoparticle labeled antibody-antigen based Lateral Flow Assay: In accordance with one or more embodiments, this technology includes but is not limited to antigens and antibodies that may be labeled with a nanoparticle or nanorod that may consist of but is not limited to gold, titanium nitride, or silica, with or without additional fluorophore dye molecules such as but not limited to rhodamine. These antigens may ^^^ compete with potential target antigens for their respective antibody. Potential target antigens may include but are not limited to bacteria surface proteins and virus proteins. The non-limiting example that can be used is a sandwich-based Lateral Flow assay to screen for the presene of B. burgdorferi. In this device, a polyclonal anti-B. burgdorferi antibody is used as the capture antibody and is immobilized as a test line on the LFA. Using a ^^^ polyclonal Ab as the capture antibody provides high affinity for the target antigen and the ability to recognize multiple epitopes on the antigen to increase chances of capturing the antigen as it flows past. A monoclonal antibody or a combination of monoclonal antibodies could also be used for the capture antibody. Targets would include OspA (which may be targeted by either or both of the signal antibody and capture antibody), Outer surface proteins ^^^ B-G, FlaB (if sample is first lysed to release this internal protein from B. burgdorferi), BBA64, or BBK32. The reason to use a monoclonal Ab as the capture Ab would be for two main reasons: monoclonal Abs are produced by a single B-cell clone in a laboratory as opposed to polyclonal Abs produced from multiple B-cell clones in different animals with varying immune responses, resulting in monoclonal Abs providing lot-to-lot consistency. ^^^ In accordance with one or more non-limiting embodiments, the signal antibody that may be used is a monoclonal anti-OspA antibody conjugated to a gold nanoparticle. Using a monoclonal Ab as the signal antibody provides the necessary specificity to the target antigen to reduce both crosslinking and false positives. Additionally, the fact that the Ab is monoclonal ensures consistency both in its conjugation to the AuNP creating the signal. ^^^ OspA was chosen as the target surface protein on B. burgdorferi due to several factors. It is one of the most researched B. burgdorferi surface proteins and is already commonly targeted in vaccine research, demonstrating its stability and immunogenic properties. As one of the major surface proteins of B. burgdorferi, it is also highly abundant on the bacteria’s surface of a tick pre-feeding. Also, OspA is relatively specific to B. burgdorferi species, reducing the ^26- ^ Docket No. L2117-7001WO likelihood of false positives due to cross-reactivity with other bacteria. It is also highly expressed in unfed ticks or ticks that have only just started a feeding, as one of its main purposes are to attach the bacteria to the tick’s midgut. Monoclonal anti-Osp C antibody may also be included as an additional signal ^^ antibody in the LFA to be used in parallel with the OspA Ab. This is because Osp C is a surface protein of B. burgdorferi that is upregulated as the tick begins to feed. If the tick is not noticed by the host and feeds for a while, OspA surface proteins begin to be downregulated in the bacteria (which could potentially result in a loss of sensitivity in the LFA), while OspC proteins begin to be upregulated. This can ensure the LFA is able to ^^^ remain able to screen for the presence of B. burgdorferi both pre-and post-tick feeding and provide a reliable result to the user. Protein BBA64 or BB0323 could also be used for the same purpose of being present post-tick feeding, plus it is present on the bacteria pre-tick feeding as well, but neither are as abundant as OspC post-feeding and have fewer commercially available antibodies. ^^^ Another non-limiting example of the LFA screening for another bacteria could be to screen for Anaplasma phagocytophillium. The capture antibody could either be a polyclonal anti-anaplasma Ab, or a monoclonal anti-MSP4 Ab. The signal Antibody conjugated to a AuNP could be a monoclonal anti-MSP4 Ab. Another non-limiting example of the LFA screening for another bacteria could be to ^^^ screen for Babesia sp., including but not limited to Babesia microti. In this LFA, the capture antibody could either be a polyclonal anti- B. microti Ab, a monoclonal anti-BmSA1 Ab, or a monoclonal anti-BmRAP-1 Ab. The signal Antibody conjugated to a AuNP could be either a monoclonal anti-BmSA1 Ab, or a monoclonal anti-BmRAP-1 Ab, whichever is not used as the capture antibody. ^^^ In accordance with one or more embodiments, lateral flow may be chosen among colorimetric assays due to the clear indication of the presence of target antigen without assistance (no color to a visible red line) versus changing from one color to another or measuring emittance using a reader. Additionally, the lateral flow assay easily allows one to see that the test worked as expected by including a control line. Additionally, the COVID-19 ^^^ pandemic caused almost ubiquitous exposure to colorimetric lateral flow assays, allowing most people to become very comfortable and proficient with using the technology. In accordance with one or more non-limiting embodiments, a signal can be detected via the LFA in about 20 minutes or less. In the non-limiting example of a B. burgdorferi LFA, the size and dimensions of the LFA could be adjusted to best screen for the target bacteria. ^27- ^ Docket No. L2117-7001WO The LFA can be competitively tuned to sense for Borrelia by having a larger pore size within the nitrocellulose membrane to allow for the larger Borrelia Burgdorferi spirochete bacteria to more easily flow through. For example, while a standard pore size is between 3-12 um, presents embodiments may involve a pore size of at least about 6-12 um. The LFA ^^ nitrocellulose membrane could have a larger pore size to better allow the B. burgdorferi (5– 40 !m in length and 0.2–0.5 !m in width) to migrate through the strip. The LFA could have a pore size that falls in the range between 8-12 um, or it could have a pore size that falls in the range of 12-20 um. Additionally, the sample buffer used with the LFA could include a surfactant including but not limited to Tween-20 to reduce bacterial clumping and improve ^^^ flow. The LFA could also forgo a conjugate pad and relegate the location of the signal antibodies to the sample buffer as shown in FIG.10 as opposed to the classic design of an LFA as shown in FIG.11. In accordance with one or more embodiments, the disclosed technology may include, but is not limited to, the use of enzymes to digest the Lyme disease causing agent B. ^^^ burgdorferi while preserving the integrity of potential diagnostic targets, including without limitation OspA. Enzymes that could be use include but are not limited to lysozyme, trypsin, and proteinase K. While B. burgdorferi has traditionally exhibited resistance to trypsin digestion, sequential or combined application of these enzymes can facilitate effective lysis. For example, lysozyme can first disrupt the outer membrane, which otherwise limits access ^^^ of trypsin to internal bacterial components. Subsequent exposure to trypsin may then degrade intracellular proteins, including periplasmic flagella that contribute to bacterial structural integrity. OspA, by contrast, naturally resists trypsin digestion, thereby becoming liberated from the bacterial matrix / outer surface in a structurally preserved form. Additionally, detergents such as Triton X-100 and Triton X-114 may be employed to ^^^ disrupt the outer surface membrane and overall bacterial structure. This can improve sample flow through the lateral flow assay and reduce issues such as damming, crosslinking, or aggregation, thereby enhancing assay sensitivity. The buffer system used for digestion and assay compatibility may include phosphate- buffered saline (PBS) or phosphate buffer. PBS mimics physiological ionic strength and can ^^^ stabilize proteins or antibodies under near-native conditions, while reducing non-specific binding. Salts within PBS may also improve flow consistency and prevent aggregation of Ab- AuNP conjugates. However, high salt concentrations can destabilize certain conjugates and promote aggregation of Ab-AuNPs. In contrast, phosphate buffer without added NaCl or KCl may reduce the risk of salt-induced aggregation and allow greater enzymatic activity, as ^28- ^ Docket No. L2117-7001WO elevated salt concentrations can inhibit trypsin. Accordingly, in some embodiments, phosphate buffer may be used preferentially as the sample buffer. A LFA could be used to screen for multiple disease vector-borne bacteria / viruses ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^molecules causing an immune response) at once. This ^^ multiplexed device could still be a sandwich-based LFA, but could have multiple capture antibodies immobilized onto the nitrocellulose strip each on their own line. The conjugate pad or the sample buffer could have multiple different signal antibodies all bound to AuNP as demonstrated in FIGS.12 and 13. As the ground vector sample interacts with the signal antibodies in either the sample buffer or the conjugate pad, the target antigens that are present ^^^ would bind to their respective signal antibodies and then would be immobilized at the correct respective capture antibody line. The antigen-antibody pairings could be to test for the prominent disease-causing bacteria / viruses in a single or multiple disease vectors in the same sample. In a non-limiting example, Borrelia burgdorferi, Anaplasma phagocytophilliu, and Babesia microti could all be tested for in a single multiplexed strip with the antibody-antigen ^^^ sandwich pairs as previously described. In accordance with one or more embodiments, the concentration of antibody bound to the NP may be optimized so that the NP is at an Optical Density (OD) visible to the human eye but not so concentrated that the required concentration of monoclonal antibodies is not economically unfeasible. An optimal balance between the NP OD and the amount of antibody ^^^ bound to the NP allows for visual inspection and appropriate detection of the bacteria in accordance with various embodiments. When creating the signal antibody by conjugating an antibody to an AuNP, the concentration ratio of antibody to AuNP could range between 20 ug / mL - 120 ug / mL to 2 – 20 OD respectively. Additionally, the concentration of signal antibody in the LFA kit sample buffer could be around 4 ug / mL with a range between 1 – 10 ^^^ ug / mL . In at least some non-limiting embodiments, a concentration of about 40 ug / mL may result in the strongest positive signal. In accordance with one or more embodiments, a unique way to detect the presence of B. burgdorferi with a lateral flow assay using both an anti-B. burgdorferi polyclonal antibody and an anti-OspA antibody (OspA is a surface protein of B. burgdorferi) is disclosed. ^^^ Polyclonal B. burgdorferi antibody targets proteins found on the outer membrane of B. burgdorferi with molecular weights of 83 kDa, 41 kDa, 34 kDa and 31 kDa and additional low MW bands protein found in / on B. burgdorferi. The 31 kDa protein that the polyclonal antibody binds to is the same OspA protein to which the monoclonal antibody binds. Monoclonal OspA antibody is a recombinant antibody (commercially available from ^29- ^ Docket No. L2117-7001WO Absolute Antibody) that targets OspA, an outer membrane surface protein of B. burgdorferi. Various other antibodies and combinations thereof can be implemented within the spirit of the invention. In accordance with one or more embodiments, the antibodies, both the NP bound- ^^ monoclonal signal antibody and the polyclonal antibodies will be on the LFA strip. In some embodiments, freeze-dried reagents that allow greater flexibility with the strip design may be implemented. A non-limiting example of how this could work is in a lateral flow assay (LFA)-based setting, a surface protein of B. burgdorferi such as but not limited to OspA may compete with ^^^ a NP-labeled OspA to bind to an anti-OspA antibody conjugated to the LFA. The fewer target OspA’s present in the sample, the more NP-labeled OspA would bind to the antibodies, resulting in a visible signal of variable strength. This could result in a binary or a semi- quantitative result where the stronger the line that appears on the LFA, the fewer target OspA (and consequently B. burgdorferi bacterium) present in the sample. ^^^ Additionally, a specific concentration of OspA could be bound to the LFA, and a specific concentration NP-labeled anti-OspA antibodies could flow down the strip with the given sample. If the sample contains OspA, those antibodies would first bind to those sample provided OspA, and leftover unbound NP-labeled antibodies bind to the LFA conjugated OspA. The fewer the leftover NP-labeled antibodies that bind to the LFA-conjugated OspA ^^^ antigens, the fainter the resultant line appears, meaning the more OspA was present in the sample. The stronger the line, the less OspA was present in the sample. Another non-limiting example of a LFA that may be used is a method involving coating a plate with a capture antibody that specifically binds to a target antigen like OspA. The sample containing the antigen is then added to the plate, allowing the antigen to bind to ^^^ the capture antibody. Subsequently, a detection antibody that either binds to OspA or another outer surface protein of B. burgdorferi, is added. This detection antibody is conjugated to an enzyme. Afterward, a substrate for the enzyme is added, producing a detectable signal, such as a color change. The intensity of the signal is measured and is proportional to the amount of antigen present in the sample. ^^^ In accordance with one or more non-limiting embodiments, the tick crushing device is a cylinder that is 3-4 inches tall, 2-3 inches in diameter. The LFA that will be inserted into the crusher is currently within the standard size range of an LFA: 3-5mm width, 50-80mm length. Current size is 4mm and 55mm length, however the length of the LFA may be increased if ^30- ^ Docket No. L2117-7001WO necessary to ensure it sufficiently extends beyond the top of the tick crusher and the positive signal line is clearly visible. In accordance with one or more embodiments, when making the “signal antibody”, an antibody (anti-OspA Ab) is generally conjugated to a gold nanoparticle AuNP. The ratio of ^^ OspA Ab to AuNP may be 40 ug / mL to 10 OD. Once made, the signal antibody is pipetted into the sample buffer solution at, for example, a 1:10 ratio (10 uL signal Ab per 100 uL sample buffer), so the final concentration of signal Ab is 4 ug OspA Ab / 1 mL Sample solution. ^^^ Nanoparticle labeled antibody-antigen based aggregation mechanism: Another non-limiting example of how antigens can be detected by NP-labeled antibodies is through an aggregation mechanism. If an antigen like B.burgdorferi is introduced into a solution of NP-labeled antibodies, the antibodies can bind to different epitopes on the antigen. This will cause the antibodies to aggregate and become cross-linked. ^^^ These aggregated larger particles could precipitate and / or cause a color change in the solution and / or cause quenching between fluorophore-associated nanoparticles that are now in close proximity to each other, with the precipitation and / or color changes indicating the presence of the antigen. ^^^ EXAMPLES Optimization of monoclonal OspA signal antibody (Ab) in LFA The purpose of this experiment was to determine the optimal concentration ratios of ^^^ Ab to 10 OD Gold nanoparticle (AuNP) during the AuNP Ab conjugation to create signal Ab. Various concentrations of the signal antibody (recombinant monoclonal OspA antibody) were tested and visual analysis of their various signal strength were evaluated. Ratios of 40, 80, and 120 ug / mL to 10 OD AuNP were tested. Specifically, concentrations of signal antibody (40, 80, 120 ug / mL) were used in a sample with B. burgdorferi. ^^^ Protocol: 1. Conjugate Ab to AuNP at varying ratios a. 40 ug / mL ^31- ^ Docket No. L2117-7001WO i. Add 100 uL of AuNP to 4 uL of Ab ii. Incubate for 30 min on a vertical rotator iii. Block the solution by adding 12.11 uL 10% BSA solution iv. Incubate for 30 more min on vertical rotator ^^ b. Repeat for 80 ug / mL and 120 i. For 80 ug / mL use 8 uL of Ab ii. For 120 ug / mL use 12 uL of Ab 2. Make Lateral Flow Assay (LFA) (dipstick test strip) a. 4x25 mm width strip of high binding Nitrocellulose Membrane from lateral flow ^^^ starter kit b. 1 uL of Biotin-labeled Rabbit Polyclonal Borrelia Antibody pipetted onto strip (Biotin-^^^^^^^^^^^^^^^^^^^^^^^^^^"^^^^^^^^^#^^^^^^^^$^^^^^^^^^^^^^^^^^^^^^^^^^ instead) (capture line) c. 1 uL of Anti-Mouse monoclonal antibody (control line) ^^^ d. Blocked with 3% BSA in PBS, washed with PBST (1X PBS and 0.5% Tween20) e. Wet edge until saturated then submerge in BSA for 15 min, wash 3 times 1 min each PBST f. Dried in vacuum desiccator for 1 hr (until dry) g. Attached onto 4x62mm Vinyl Backing Card, overlayed with ~4x15mm Wicking Pad ^^^ with ~ 2-5mm overlay to ensure proper wicking contact 3. Create sample solution with positive Borrelia sample and signal Ab-AuNP conjugate Component Volume . i. 3 samples each with signal Ab concentration of 40, 80, 120 ug / mL ii. 1 “negative” sample that did not contain any Borrelia in the sample ^32- ^ Docket No. L2117-7001WO The related data is presented in FIG.14 reflecting the varying signal antibody concentrations and the LFA positive signal strength. The red circular dot reflects a positive signal for the presence of B. Burgdorferi. The red line at the bottom of each LFA strip ^^ matches the height where the sample solution surface met the LFA, and is indicative of Borrelia-signal Ab-AuNP conjugates that are too large to flow up the strip. The 40 ug / mL Signal Ab to 10OD AuNP resulted in the strongest positive signal. Confirming positive correlation between B. burgdorferi concentration and signal strength ^^^ An experiment was conducted to confirm that with diluted concentrations of B. burgdorferi in a sample, the LFA gives a less vibrant positive signal, showing a positive qualitative correlation between B. burgdorferi concentration and signal strength. The purpose of the experiment was to verify that there is a direct correlation between B. burgdorferi concentration and LFA signal strength in the test strip. LFA is screening for the ^^^ presence of B. burgdorferi using a polyclonal Anti-B. burgdorferi antibody as a capture antibody, and Anti-OspA antibody labeled with AuNP as a signal antibody. If the sandwich assay is correctly capturing B. burgdorferi using the given capture and signal Abs, decreasing concentrations of B. burgdorferi in the sample should correlate to a decreasing positive signal presented on the LFA. A series of dilutions from 1:1 to 1:1000 dilutions were made, and LFA ^^^ copies were inserted into each one. One factor that may affect this experiment is the crosslinking that could be occurring at the point of contact between LFA and sample solution, where crosslinked Borrelia-labeled signal Ab is too large to move up the strip, potentially affecting the signal strength for each strip differently. Protocol: ^^^ 1. LFA creation a. 1 uL of Biotin-labeled Rabbit Polyclonal Anti-Borrelia Antibody pipetted onto strip (Biotin-^^^^^^^^^^^^^^^^^^^^^^^^^^"^^^^^^^^^^^^^-borrelia Polyclonal Ab recommended instead) (capture line) b. 1 uL of Anti-Mouse monoclonal antibody (control line) ^^^ c. Blocked with 3% BSA in PBS, washed with PBST (1X PBS and 0.5% Tween20) ^33- ^ Docket No. L2117-7001WO d. Wet edge until saturated then submerge in BSA for 15 min, wash 3 times 1 min each PBST e. Dried in vacuum desiccator for 1 hr (until dry) f. Attached onto 4x62mm Vinyl Backing Card, overlayed with ~4x15mm Wicking Pad ^^ with ~ 2-5mm overlay to ensure proper wicking contact 2. OspA monoclonal Ab and AuNP conjugation per attached AuNP-Ab conjugation SOP a. Briefly: i. Dilute AuNP to OD of 10OD ii. Pipette 4 uL antibody per 100 uL AuNP 10 OD ^^^ iii. Incubate on vertical rotator for 30 min iv. Block with 1% BSA v. Incubate on vertical rotator for 30 min 3. Testing dilutions of B. burgdorferi ^$ Current optimized standard sample solution:^^ ^^^ ^ Component Volume 4. Test samples with B. burgdorferi dilutions: a. 1:120 uL B. burgdorferi b. 1:210 uL B. burgdorferi, 10 uL H2O ^^^ c. 1:54 uL B. burgdorferi, 16 uL H2O d. 1:104 uL B. burgdorferi, 36 uL H2O ^34- ^ Docket No. L2117-7001WO e. 1:10012 uL of 1:10 soln, 108 uL H2O f. 1:1K 12 uL of 1:100 soln, 108 uL H2O 5. Make samples that substitute the 20 uL “Heat-treated B. burgdorferi sample (positive control)” ^^ with 20uL of each dilution listed above + 1 negative control sample with no B. burgdorferi 6. Incubate for 10 minutes 7. Transfer to well plate and insert LFA, measure at 20 min. Results are presented in FIG.15. From left to right: 1 : 1, 2, 5, 10, 100, 1000 dilutions ^^^ of B. burgdorferi sample. Right most well at H7 is negative control. It was observed as the samples become more diluted with B. burgdorferi, the signal strength becomes weaker. There is a clear qualitative result showing a positive correlation between B. burgdorferi concentration and positive signal strength. Also of note, the “bottom red line” is indicative that either B. burgdorferi - signal Ab crosslinking is occurring, or most ^^^ B. burgdorferi - signal Ab conjugates are too large to migrate up the strip. AuNP-OspA Ab signal antibody conjugation Gold nanoparticle (AuNP) labeled Anti-OspA antibodies are not readily commercially available. The following is a non-limiting SOP to prepare AuNP- labeled Anti-OspA Ab for use in a LFA with a concentration ratio of 40 ug / mL OspA Ab per 10 OD AuNP. Previous ^^^ experiments established that pH 8.0 is the optimal pH to conjugate Ab to AuNP, and previous experiment established the 40ug / mL Ab to 10 OD AuNP was the optimal concentration ratio for use in an LFA. Protocol: 1. Dilute stock 20OD Au Soln to result in 200 uL 10 OD: ^^^ a. Materials: i. 20 mM citrate pH 5.0 ii. 0.2 mM citrate solution iii. Au soln 20 OD b. Separately, prepare a stock solution of 0.2 mM citrate by diluting the 20 mM ^^^ citrate 1 / 100 in diH2O ^35- ^ Docket No. L2117-7001WO c. In new eppendorf tube, add 100 uL Au soln OD 20 d. Add 1 uL 20 mM citrate buffer to the eppendorf tube (results in 20 OD Au in 0.2 mM citrate solution) e. Add 99 uL of 0.2 mM citrate soln to achieve 200 uL of 10 OD Au. ^^ 2. Conjugate Ab to AuNP a. Materials: i. 4uL anti OspA mouse monoclonal Ab ii. 5 uL 100 mM phosphate buffer 8.0 iii. 100 uL 10 OD Au soln ^^^ iv. 10% BSA in H2O b. In new eppendorf tube, add 5 uL of 100 mM phosphate buffer pH 8.0 c. Then 4 uL Ab d. Then add 100 uL of 10 OD AuNP aliquot e. Tap with finger to mix, then put on vertical rotator to incubate / mix for the next ^^^ 30 min. f. After incubating 30 min, add 10.9 uL 10%BSA in H2O to result in 1% BSA in the soln g. Incubate for an additional 30 min on vertical rotator Results in ~ 100 uL AuNP labeled OspA Ab signal antibody ready for use in a LFA. ^^^ ^36- ^ Docket No. L2117-7001WO The phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. As used herein, the term “plurality” refers to two or more items or components. The terms “comprising,” “including,” “carrying,” “having,” “containing,” and “involving,” whether in the written description or the claims and the like, ^^ are open-ended terms, i.e., to mean “including but not limited to.” Thus, the use of such terms is meant to encompass the items listed thereafter, and equivalents thereof, as well as additional items. Only the transitional phrases “consisting of” and “consisting essentially of,” are closed or semi-closed transitional phrases, respectively, with respect to the claims. Use of ordinal terms such as “first,” “second,” “third,” and the like in the claims to modify a claim ^^^ element does not by itself connote any priority, precedence, or order of one claim element over another or the temporal order in which acts of a method are performed, but are used merely as labels to distinguish one claim element having a certain name from another element having a same name (but for use of the ordinal term) to distinguish the claim elements. Having thus described several aspects of at least one embodiment, it is to be ^^^ appreciated various alterations, modifications, and improvements will readily occur to those skilled in the art. Any feature described in any embodiment may be included in or substituted for any feature of any other embodiment. Such alterations, modifications, and improvements are intended to be part of this disclosure and are intended to be within the scope of the invention. Accordingly, the foregoing description and drawings are by way of example only. ^^^ Those skilled in the art should appreciate that the parameters and configurations described herein are exemplary and that actual parameters and / or configurations will depend on the specific application in which the disclosed methods and materials are used. Those skilled in the art should also recognize or be able to ascertain, using no more than routine experimentation, equivalents to the specific embodiments disclosed. ^ ^^^ ^37- ^

Claims

Docket No. L2117-7001WO CLAIMS What is claimed is:

1. A device for processing a disease vector for screening, comprising: a receptacle defining a crushing recess constructed and arranged to receive a disease ^^ ^^^^^^^^^^^^^^^^^^^^^^ a cap configured to create a self-contained system and not reopen when securably mated with the receptacle while still being rotatable relative to the receptacle to facilitate processing of the disease vector within the self-contained ^^^^^^^^^^^ a crushing element extending from the cap into the receptacle and configured to ^^^ engage with the crushing recess via rotation of the cap to process the disease vector.

2. The device of any of the preceding claims, wherein processing the disease vector comprises at least partially fragmenting the disease vector. ^^^ 3. The device of any of the preceding claims, wherein the self-contained system is configured to exert a mechanical force on the disease vector.

4. The device of any of the preceding claims, wherein the self-contained system is configured to exert mechanical force on the disease vector in two axis. ^^^ 5. The device of any of the preceding claims, wherein at least one of the crushing recess and the crushing element define a surface roughness to facilitate processing of the disease vector.

6. The device of any of the preceding claims, wherein the crushing element comprises a ^^^ crushing head connected to a crushing implement.

7. The device of any of the preceding claims, wherein the crushing element comprises one or more contact portions to facilitate processing of the disease vector. ^^^ 8. The device of any of the preceding claims, wherein the crushing implement is guided by cam followers engaged with a barrel cam track.

9. The device of any of the preceding claims, wherein the crushing surfaces comprise annular ribs with saw-cut profiles. ^38- ^Docket No. L2117-7001WO 10. The device of any of the preceding claims, further comprising openings between the grinding chamber and a sample chamber to permit post-crushing mixing with a reagent fluid. ^^ 11. The device of any of the preceding claims, wherein the cap includes a one-way locking mechanism and a diagnostic test strip insertion port.

12. The device of any of the preceding claims, wherein the receptacle and cap are ergonomically contoured and textured for one-handed operation. ^^^ 13. The device of any of the preceding claims, wherein the receptacle is further configured to receive fluid to facilitate screening of the processed disease vector.

14. The device of any of the preceding claims, wherein the device further comprises a fluid ^^^ chamber.

15. The device of any of the preceding claims, wherein the fluid chamber is contained within the lid or receptacle. ^^^ 16. The device of any of the preceding claims, wherein the device further comprises a separation mechanism to facilitate downstream disease vector screening.

17. The device of any of the preceding claims, wherein the device is connectable to an assay device. ^^^ 18. The device of any of the preceding claims, wherein the assay device does not involve a DNA-based assay.

19. The device of any of the preceding claims, wherein the device is configured to receive an ^^^ assay test strip to facilitate disease vector screening.

20. The device of any of the preceding claims, wherein screening the disease vector involves a fluoroimmunoassay. ^39- ^Docket No. L2117-7001WO 21. The device of any of the preceding claims, wherein the assay device is a colorimetric assay. 22.The device of any of the preceding claims, wherein screening the disease vector involves ^^ nanoparticle (NP)-labeled antibody and / or antigen-based sensing.

23. The device of any of the preceding claims, wherein the assay device uses both an anti-B. burgdorferi polyclonal antibody and an anti-OspA antibody. ^^^ 24. The device of any of the preceding claims wherein the concentration of signal antibody is about 40 ug / mL.

25. The device of any of the preceding claims, wherein the device is a tick screening device intended for at-home use. ^^^ 26. A lateral flow assay (LFA) for screening a disease vector, comprising: an anti-B. burgdorferi polyclonal antibody and an anti-OspA antibody.

27. A kit, comprising: ^^^ the device for processing a disease vector of any of the preceding claims.

28. The kit of any of the preceding claims, further comprising a source of a buffer solution.

29. The kit of any of the preceding claims, further comprising a source of labeled antibodies. ^^^ 30. The kit of any of the preceding claims, wherein the source of labeled antibodies comprises a sealed dropper of a buffer solution containing the labeled antibodies.

31. The kit of any of the preceding claims, further comprising an assay device. ^^^ 32. The kit of any of the preceding claims, wherein the assay device is an antibody and / or antigen-based sensing assay.

33. The kit of any of the preceding claims, wherein the assay device is a fluoroimmunoassay. ^40- ^Docket No. L2117-7001WO 34. The kit of any of the preceding claims, wherein the assay device is a colorimetric assay device. ^^ 35. The kit of any of the preceding claims, wherein the assay device is a lateral flow assay (LFA).

36. The kit of any of the preceding claims, wherein the assay device involves an LFA test strip. ^^^ 37. The kit of any of the preceding claims, wherein the assay device uses both an anti-B. burgdorferi polyclonal antibody and an anti-OspA antibody.

38. The kit of any of the preceding claims, wherein the assay device uses an anti-B. ^^^ burgdorferi polyclonal antibody, an anti-OspA antibody, and an anti-OspC antibody.

39. The kit of any of the preceding claims, wherein the concentration of signal antibody- AuNP present in the LFA sample buffer is about 4 ug / mL. ^^^ 40. The kit of any of the preceding claims, wherein the signal antibody’s antibody to AuNP conjugation ratio is about 40 ug / mL per 10 OD.

41. The kit of any of the preceding claims, wherein the assay device screens for multiple vector-borne diseases in a single sample using a unique capture-signal antibody sandwich ^^^ combination for each bacteria tested.

42. The kit of any of the preceding claims, wherein the LFA test strip is defined by an average pore size of at least about 6-12 uM. ^^^ 43. The kit of any of the preceding claims, wherein the assay screens for a protein associated with Anaplasma phagocytophillium or Babesia microti.

44. The kit of any of the preceding claims, wherein the LFA test strip does not include a conjugate pad on which the signal antibody is immobilized. ^41- ^Docket No. L2117-7001WO 45. The kit of any of the preceding claims, wherein the signal antibody is present in a buffer solution. ^^ 46. The kit of any of the preceding claims, wherein the assay is an aggregation-based assay.

47. The kit of any of the preceding claims, further comprising a base configured to stabilize the receptacle during processing of the disease vector. ^^^ 48. The kit of any of the preceding claims, wherein the kit is intended for at-home use.

49. The kit of any of the preceding claims, wherein the kit is a Lyme disease screening kit.

50. A method of screening a disease vector, comprising: ^^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^%^^^^^^^^^^^^^^^^^^^^^^^^^^^ subjecting the solution to an assay to assess the presence or absence of a disease-causing pathogen or molecule causing an immune response.

51. The method of any of the preceding claims, wherein processing the disease vector ^^^ involves applying mechanical force across two axis.

52. The method of any of the preceding claims, wherein the assay is not a DNA-based assay.

53. The method of any of the preceding claims, wherein the assay screens for a target protein ^^^ associated with the disease-causing pathogen or molecule causing an immune response.

54. The method of any of the preceding claims, wherein the disease-causing pathogen or molecule causing an immune response is associated with Lyme disease, babesiosis, ehrlichiosis, Rocky Mountain Spotted Fever, anaplasmosis, Southern Tick-Associated Rash ^^^ Illness, Tick-Borne Relapsing Fever and tularemia.

55. The method of any of the preceding claims, wherein the target protein is associated with a Lyme disease-causing bacteria. ^42- ^Docket No. L2117-7001WO 56. The method of any of the preceding claims, wherein the target protein is associated with B. burgdorferi.

57. The method of any of the preceding claims, wherein the assay targets a B. burgdorferi ^^ protein.

58. The method of any of the preceding claims, wherein the assay targets outer surface protein A (OspA). ^^^ 59. The method of any of the preceding claims, wherein the assay targets a tick salivary gland protein.

60. The method of any of the preceding claims, wherein the assay is an antibody and / or antigen-based sensing assay. ^^^ 61. The method of any of the preceding claims, wherein the assay is a lateral flow assay (LFA).

62. The method of any of the preceding claims, wherein the assay is an aggregation-based ^^^ assay.

63. The method of any of the preceding claims, wherein the assay further screens for a second target protein associated with a second disease-causing bacteria. ^^^ 64. Any lateral flow assay (LFA) for screening a disease vector as described herein.

65. The LFA of any of the preceding claims, wherein the LFA screens for a target protein associated with a Lyme disease-causing bacteria. ^43- ^

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