200 results about "Canine parvovirus" patented technology

A rapid test apparatus, system, and method of use utilizing lateral flow immunoassay (LFIA) detection of a selected ligand in a liquid sample from a body fluid such as saliva in a pet in which antibodies and their complimentary antigens are used with detection-nanoparticles to provide a visual or measurable end point indicator in which the method measures the exposure to viruses in the canine from Canine Parvovirus (CPV) and / or Canine Distemper virus (CDV).

The purpose of the invention is to disclose a porcine circovirus, porcine parvovirus and porcine reproductive and respiratory syndrome virus triplex virus-like particle vaccine and its preparation method. The triple virus-like particle vaccine (Triple VLP vaccine) of the invention contains VLP which is composed of PCV-2 major structural protein CAP protein, PPV VP2 protein epitope and PRRSV Gp5 protein epitope. It is proved by experiment that the vaccine can stimulate good double cellular and humoral immune response. It is shown by pharmacodynamic test that after immunization of different animal groups, the vaccine of injection, nose drops and water forms prepared by VLP antigen formed by the method with or without adjuvants can safely and effectively prevent the infection of PCV-2, PPV and PRRSV. The invention provides an ideal vaccine for the security of sows, piglets and fattening pigs to effectively prevent mixed infection of PCV-2, PPV and PRRSV.

The invention provides a canine parvovirus strain CPV-YH and applications thereof, and belongs to the technical field of microbes. The preservation number of the provided canine parvovirus strain CPV-YH is CGMCC No.12990. The invention also provides applications of the canine parvovirus strain CPV-YH in the preparation of canine parvovirus vaccines and canine parvovirus diagnostic reagents. Based on the canine parvovirus strain CPV-YH, the invention also provides a vaccine composition for preventing and / or treating diseases caused by canine parvovirus and a reagent for detecting a canine parvovirus strain and / or diagnosing diseases caused by canine parvovirus. A novel content and direction are provided for the research on the variation of epidemic virus; a novel strain is provided for the canine parvovirus vaccine development; and a high quality raw material and choice are provided for the vaccine development.

The invention discloses a bivalent live vaccine against canine distemper and parvovirus diseases, and a preparation method thereof. The bivalent live vaccine contains an attenuated anti-canine distemper vaccine strain with a microbial accession number of CGMCC No. 10980 and an attenuated anti-parvovirus distemper vaccine strain with a microbial accession number of CGMCC No. 7408. According to the invention, a canine distemper virus and a canine parvovirus separated from wild viruses of diseased Chinese dogs and having undergone attenuation via domestication are used for preparation of a bivalent vaccine together, and a freeze-drying protective agent is added so as to prepare the bivalent live vaccine against canine distemper and parvovirus diseases. The bivalent live vaccine is reliable in security, good in immune effect, strong in targeting performance, suitable for prevention of diseased caused by diseased Chinese dogs and applicable to prevention of canine distemper and parvovirus diseases.

The invention discloses a mink distemper-canine parvovirus enteritis bivalent vaccine and its preparation method and use and belongs to the technical field of bioengineering. The mink distemper-canine parvovirus enteritis bivalent vaccine mainly comprises a mink distemper live vaccine, a mink canine parvovirus enteritis inactivated vaccine, a vaccine adjuvant and a protective agent. Aiming at one object, a ratio of the mink distemper live vaccine to the mink canine parvovirus enteritis inactivated vaccine is 1: 1. The invention also provides a preparation method and a use method of the mink distemper-canine parvovirus enteritis bivalent vaccine. The mink distemper-canine parvovirus enteritis bivalent vaccine is safe, has high efficiency, can save labor and can reduce a culture cost.

The invention provides canine parvovirus-like particles. The VP2 gene of canine parvovirus is optimized according to the preferred codons of insect cells, the optimized VP2 gene is connected with an insect expression vector, and the canine parvovirus-like particles are produced by use of a rhabdovirus / insect cell expression system. The canine parvovirus-like particles are combined with a preservative and an adjuvant to prepare a canine parvovirus enteritis vaccine. By adopting the canine parvovirus-like particles provided by the invention, the prepared canine parvovirus enteritis vaccine has the advantages of high output, low production cost, high immunogenicity, good safety, convenience in large-scale production and the like, and has good immunization and prevention effects on canine parvovirus enteritis.

The invention discloses a method for producing a curative canine parvovirus virus monoclonal antibody by using a bioreactor.

The invention belongs to the technical field of animal epidemic disease detection and quarantine, and discloses a canine distemper virus (CDV) and canine parvovirus (CPV) duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and a detection method thereof. The detection kit comprises a pair of specific primers designed based on CDVH genes, a TaqMan-MGB probe with the 5' end marked as FAM, a pair of specific primers designed based on CPVVP2 genes and a TaqMan-MGB probe with the 5' end marked as VIC. The detection method comprises the following steps: designing two pairs of primer sequences and two TaqMan-MGB marked probe sequences, and performing fluorescent labeling; preparing total RNA (ribonucleic acid) and total DNA (deoxyribonucleic acid); performing reverse transcription; performing fluorescent PCR amplification; performing result judgment. According to the detection method, the nucleic acids of a CDV and a CPV can be detected at the same time and the CDV / CPV can be quickly, specifically and sensitively identified at the same time. The detection kit is of great significance on prevention and control of CDV and CPV diseases.

The invention discloses a two-color fluorescence PCR primer, probe and method for quickly distinguishing canine parvovirus vaccine strain from wild strain. The real-time fluorescence PCR technique and the melting curve analysis technique are combined, and canine parvovirus vaccine strain and wild strain are identified according to the peak pattern of a melting curve and Tm value difference; operation is easy, and identification and detection of three kinds of wild strain with different genotypes and two kinds of vaccine strain can be achieved through one reaction; detection speed is high, and flux is high; the whole operation process can be finished within 3 h, cell cultivation of viruses is not needed, and time for identification and detection of three kinds of wild strain with different genotypes and two kinds of vaccine strain is shortened greatly; accuracy, specificity and repeatability are high, accurate, quick and high-flux analysis can be achieved, and application and popularization in clinical practice can be achieved easily.

The invention discloses a production method of a canine parvovirus inactivated vaccine by utilizing a bioreactor. The production method comprises the following steps: (1) culturing a vaccine producing cell by applying a microcarrier and chip carrier system of the bioreactor; (2) vaccinating a canine parvovirus and performing virus multiplication culture; (3) harvesting virus culture fluid; and (4) inactivating the virus fluid with BEI (binary ethyleneimine), and adding adjuvant to prepare the vaccine. The canine parvovirus inactivated vaccine has the advantages of high density of cultured cell, high virus titer, uniform and stable quality, controllable process and high production efficiency, and the defects of large difference among product batches, low antigen content and low production efficiency in a conventional spinner bottle or chick embryo production process can be overcome.

The invention relates to a yolk antibody injection against canicola fever and canine parvovirus and a preparation method thereof. The yolk antibody injection against canicola fever and canine parvovirus is extracted and prepared by the collected yolk antibody of the immunized laying hens which are immunized by the bivalent vaccine of the canicola fever and canine parvovirus. The yolk antibody injection against canicola fever and canine parvovirus prevents the diseases by improving the antibody level of canine bodies to the canicola fever and the canine parvovirus. The yolk antibody injection against canicola fever and canine parvovirus and a preparation method thereof have the beneficial effect that the anti-canicola fever and canine parvovirus disease egg yolk antibody injection is provided with faster effect in the onset of canicola fever and canine parvovirus diseases. The yolk antibody injection against canicola fever and canine parvovirus and the preparation method thereof has the advantages of obvious efficacy, low cost, fast effect, no side effects, no generated drug resistance, simple preparation, wide raw material resource, convenient use and easy application.

The invention discloses a canine parvovirus virus-like particle, and a preparation method and a use thereof. The canine parvovirus virus-like particles provided by the invention are formed by VP2 protein; the gene sequence of the canine parvovirus virus-like particles is SEQ ID No 1. The preparation method of the canine parvovirus virus-like particles provided by the invention comprises the following steps of: by taking the DNA (Deoxyribose Nucleic Acid) of the canine parvovirus as a template, amplifying the DNA with SEQ ID No 2 and SEQ ID No 3 as primers to obtain an amplification product, inserting the amplification product into a carrier pSMK to obtain a recombinant plasmid pSMK-VP2, transforming the pSMK-VP2 to an escherichia coli BL21-CondonPlus (DE3)-RIL line, culturing in an LB (Luria-Bertani) culture medium and expressing a recombinant protein through induction by using IPTG (Isopropyl-beta-D-Thio Galactopyranoside), and then purifying the recombinant protein and carrying out in-vitro assembly to obtain the canine parvovirus virus-like particles.

The invention discloses a PCR-HRM primer and method for quickly distinguishing canine parvoviruses of different genotypes. The method comprises the following steps: extracting virus DNA (deoxyribonucleic acid) from a sample; by taking the virus DNA as a template, using a designed specific primer pair and fluorescent saturated dye to perform amplification reaction, thus obtaining an amplification product; and finally, performing HRM analysis on the amplification product, thus determining the genotype of a canine parvovirus. According to the invention, the method is simple to operate, only the fluorescent saturated dye needs to be added before PCR reaction, and the method is high in detection speed and high in flux; the whole operation process only costs 3 hours, and cell culture of viruses is not needed, thus greatly shortening the time required for typing; the expense is low, specific probes are not needed, and the cost of the saturated dye for each sample is 1.6RMB; and the accuracy is high, the specificity and repetitiveness are favorable, and analysis can be accurately and quickly performed at high flux, thereby ensuring that the invention is beneficial to popularization and application in clinical practice.

The invention discloses a canine parvovirus single-domain antibody, and a preparation method and an application thereof. The canine parvovirus single-domain antibody has a nucleotide sequence as shown in SEQIDNO.1. The preparation method comprises the steps: immunizing an alpaca by a canine parvovirus inactivated vaccine, and taking peripheral blood to obtain alpaca anti-canine parvovirus serum; extracting RNA in the peripheral blood, and carrying out reverse transcription to generate cDNA; utilizing specific primers for amplification, and purifying a VHH fragment; connecting the VHH fragment with a phagemid vector, transforming TG1, and constructing a canine parvovirus single-domain antibody library; and enriching the antibody library by utilizing a canine parvovirus structural protein VP2, introducing into an expression bacterial strain E.coil HB2151, and utilizing IPTG to induce the canine parvovirus single-domain antibody VHH having the biological activity to be expressed. The prepared canine parvovirus single-domain antibody has the detection activity of 15 [mu]g / ml and the detection sensitivity of 250 ng / ml, and can be used as drugs applied for treating dogs with canine parvovirus disease.

The invention discloses a canine distemper and canine parvovirus disease bivalent vaccine and a preparation method thereof. The bivalent vaccine is prepared by compounding a canine distemper vaccine and a canine parvovirus disease vaccine according to a volume ratio of 1:1, and then adding a freeze-dried protective agent and a multi-bacteria inhibitor into the mixed vaccine. The provided canine distemper and canine parvovirus disease bivalent vaccine is prepared by compounding a canine distemper vaccine and a canine parvovirus disease vaccine, at the same time a freeze-dried protective agent and a multi-bacteria inhibitor are added to enhance the effect, and the bivalent vaccine has the advantages of stable quality, long immunity time, few adverse reactions, safety, and reliability. At the same time, the bivalent vaccine can simultaneously effectively control the canine distemper virus and canine parvovirus, and has a prominent immunity effect. The application dosage is equal to that of a mono-valent vaccine, and the effect of the bivalent vaccine is better than the total effect of two mono-valent vaccines. The vaccination procedure is simplified, the frequency of stress reactions of vaccinated animals is reduced, and one vaccine can prevent two diseases.

The invention discloses a preparation method of an anti-canine parvovirus protein VP2 specific IgY, which comprises the following steps: (1) designing a pair of primers according to CPV-VP2 gene sequence, carrying out PCR (polymerase chain reaction) amplification on the CPV-VP2 gene, connecting to a pMD18-T vector, transforming DH5alpha competent cell, carrying out blue-white screening, extracting the plasmid, carrying out digest enzyme digestion ion analysis, carrying out positive plasmid sequencing, and carrying out comparative analysis on the sequencing result; (2) expression and purification of VP2 protein in Escherichia coli: carrying out BamH I and Xho I double-enzyme digestion on the pMD18-T-VP2 and pET-32a,connecting to the target segment, constructing the pET-32a-VP2 expression vector, transforming Bal21(DE3)pLysS competent bacteria, carrying out enzyme digestion and PCR identification, optimizing the IPTG (isopropyl-beta-D-thiogalactopyranoside) induction concentration and time, carrying out mass induction expression, and purifying the recombinant protein by using a Ni<+> affinity column; and (3) preparation of anti-VP2-IgY antibody: immunizing a laying hen by using the purified VP2 protein, extracting the specific IgY antibody by using PEG 6000, and carrying out SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) analysis. The anti-VP2-IgY antibody extracted by the method can be well combined with the VP2 protein to carry out non-cross reaction on the degradation segment.

The invention discloses a porcine parvovirus L strain and use thereof in preparation of porcine parvovirus inactivated vaccines. In the invention, microbial preservation number of the separated strain is CGMCC No. 3352. The strain is highly homologous with an NADL-2 strain and a China strain and keeps high reproduction, and the viruses are stable. Oil adjuvant is added into antigen liquid which inactivates the viruses by using binary ethylenimine to prepare the bidirectional oil-emulsion inactivated vaccines; and the prepared inactivated vaccines are applied in a vaccine inoculation experiment of 10,276 pigs (most of which are first farrowing sows), and in the experiment, an immune dosage of 2 millimeters is given to each pig through muscle injection, and each time of inactivated vaccine inoculation can obtain an immune period of over 6 months. Under the conditions that a 10 millimeter overdose vaccine is injected into a pig and that single dose inoculation is repeatedly carried out for three times, no abnormal reaction happens, which proves that the inactivated vaccines have high immunogenicity and safety.

The invention discloses a set of multiplex polymerase chain reaction (PCR) detection primers for detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus. The primers comprise three pairs of primers, wherein sequences of primers for detecting canine distemper virus are shown as SEQ ID NO:1 and SEQ ID NO:2; sequences of primers for detecting canine parvovirus are shown as SEQ ID NO:3 and SEQ ID NO:4; and sequences of primers for detecting type I canine adenovirus and type II canine adenovirus are shown as SEQ ID NO:5 and SEQ ID NO:6. The primers can simultaneously detect canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus through a multiplex PCR method, and have the advantages of high specificity, high sensitivity, high repeatability and the like.

The invention discloses a canine parvovirus colloidal gold immunochromatography test strip and a preparation method. The test strip comprises a sample absorbing area, a gold label probe area, an immobilized antibody area, a water absorbing area and a bottom plate, wherein the sample absorbing area, the gold label probe area, the immobilized antibody area and the water absorbing area are sequentially paved on the bottom plate and partially overlapped each other; the gold label probe area is coated with a colloidal gold labeled anti-canine parvovirus monoclonal antibody F1; the immobilized antibody area is sequentially provided with a test line and a control line, the test line is coated with an anti-canine parvovirus monoclonal antibody B6, and the control line is coated goat anti-mouse IgG (immunoglobulin G). According to the preparation method, a glass fiber membrane, a polyester membrane coated with the anti-canine parvovirus monoclonal antibody F1, a nitrocellulose membrane coated with the test line and the control line, and water absorbing filter paper are assembled onto a polyethylene plate to obtain the canine parvovirus colloidal gold immunochromatography test strip. The test strip is fast to test, high in accuracy rate, high in specificity, and simple and convenient to carry and operate.

The invention discloses a recombinant rabies virus rHEP-Flury(dG-VP2) carrying two genes G and a gene VP2. According to the recombinant virus, a rabies virus HEP-Flury strain serves as a framework, the VP2 gene shown as SEQ ID NO.1 is inserted into HEP-Flury, an additional gene G of the rabies virus is inserted, and the recombinant plasmid pHEP-Flury(dG-VP2) carrying the two genes G and the gene VP2 is obtained; finally, rescuing screening is carried out, and the recombinant rabies virus rHEP-Flury(dG-VP2) is obtained. The recombinant rabies virus rHEP-Flury(dG-VP2) carries the two genes G and expresses VP2 protein, a high rabies virus resisting antibody and canine parvovirus VP2 resisting antibody level can be generated through immune induction, the cost of vaccines for dogs can be reduced, and very good application and popularization prospects are achieved.

The invention discloses a PCR-HRM primer and a method for rapidly distinguishing a wild strain and a vaccine strain of canine parvovirus. The method comprises the following steps of: extracting a virus DNA from a sample as a template, carrying out PCR amplification by utilizing designed two pairs of specific primers and fluorescent saturated dye, carrying out HRM analysis on the detected sample respectively with a wild strain standard sample and a vaccine strain standard sample as contrast, and determining the type of the canine parvovirus. With the adoption of the method, the operation is simple, and only is the fluorescent saturated dye added before the PCR; the detection speed is high, the flux is high, the whole operation process only needs 3 hours, and cell culture of viruses is not needed, and thus the time needed by distinguishing detection is greatly shortened; the cost is low, a specific probe is not needed, and the fluorescent saturated dye is cheap and easily obtained; as the accuracy is high, the specificity and the repeatability are good, and analysis can be accurately and rapidly carried out with high flux, the method is beneficial to popularizing and applying in clinical practices.

The invention relates to an efficient-secretion canine parvovirus (CPV) resistant monoclonal antibody hybridoma cell A135 strain and belongs to the technical field of biology. The hybridoma cell A135 strain selected from the secretion CPV resistant monoclonal antibody hybridoma cell bank is excellent in biological performance and is injected with ascetic fluid generated in a BALB / C mouse peritoneal, so that neutralizing titer is up to 1010. In addition, the hybridoma cell A135 strain has moderate neutralizing capacity for various CPV subtype virus strain such as CPV-2a, CPV-2b, CPV-2c (a) and CPV-2c (b) and CPV strains from foxes and raccoon dogs, is wide in anti-CPV strain range and is used for clinical treatment of attacked dogs with CPV diseases, and effective rate is up to 100%.

The invention discloses a reagent kit for quickly detecting nucleic acid of canine parvovirus (CPV). The reagent kit comprises lysate, diluent, lyophozyme tubes with primers, detection liquid and test strips. The reagent kit has the advantages that normal-temperature nucleic acid amplification technologies are adopted, different types of enzymatic reaction can be carried out by the aid of DNA (deoxyribonucleic acid) helicase, single-stranded DNA binding proteins, DNA polymerase and the like, and accordingly to-be-detected DNA can be amplified by several millions of times under the condition of the constant temperatures of 37-45 DEG C within 10-30 minutes; the reagent kit is matched with immune colloidal gold chromatographic detection technologies, and accordingly the to-be-detected DNA can be quickly verified; the reagent kit is easy to operate, short in time and quite suitable for quickly diagnosing infectious diseases of pets and has low requirements on instruments.

The invention relates to the technical field of biology, in particular to an LAMP kit for detecting a canine parvovirus. The kit comprises six LAMP primers, wherein the LAMP primers and LAMP reaction liquid form a detecting system together. The kit can rapidly and sensitively detect various current canine parvovirus strains, has simple operation, low cost and favorable specificity, is quite suitable for field detection of export quarantine, food sanitation and livestock farms and is easy for generalization and application in a large range, and a reaction result is easy to observe.

The invention discloses a two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses). A detection method comprises the following steps: firstly, pre-treating a sample by using a nucleic acid extraction solution of the kit and extracting nucleic acid; then detecting the nucleic acids of the CDV and CPV simultaneously in a reaction system by utilizing the two-color fluorogenic quantitative PCR detection kit, and detecting whether the canine distemper and canine parvo viruses exist in the sample to be detected or not by one step; and quantifying a virus copy number. The detection method is high in detection efficiency and good in specificity and sensibility; the aims of clinical detection and epidemic situation control of animal epidemic diseases can be realized.

The invention relates to a traditional Chinese medicine injection for treating canine parvovirus disease, wherein crude drugs of the injection consist of following components: 100 parts by weight (pbw) to 500pbw of Chinese pulsatilla root, 100pbw to 300pbw of indigowoad leaves, 100pbw to 400pbw of honeysuckle flowers, 10pbw to 100pbw of rhubarb, 100pbw to 200pbw of flos sophorae, 100pbw to 200pbw of rhizoma coptidis, 100pbw to 300pbw of radix astragali, 100pbw to 300pbw of dark plum fruits, 100pbw to 200pbw of radix curcumae, 100pbw to 300pbw of myrobalam, 100pbw to 300pbw of hairyvein agrimonia herb and bud and 100pbw to 200pbw of rhizoma pinelliae processed with gingers; the invention also discloses a preparation method of the injection, which comprises the steps of: conducting steam distillation extraction on the Chinese pulsatilla root and the radix curcumae, conducting alcohol extraction on the hairyvein agrimonia herb and the rhizoma pinelliae, and conducting water-alcohol extraction on the indigowoad leaves, the honeysuckle flowers, the rhubarb, the flos sophorae, the rhizoma coptidis, the radix astragali, the dark plum fruits and the myrobalam, thereby obtaining the injection. The injection can achieve the purpose of treating the canine parvovirus disease by clearing away heat and toxic material, dispelling heat and relaxing bowels, relieving diarrhea with astringents, cooling blood to stop blood, calming the adverse-rising energy and arresting vomiting. The injection is the traditional Chinese medicine injection prepared by extracting and refining traditional Chinese medicinal materials, thus having no toxicity or side effect on the canine, with quick effect and high cure rate after injection.

The invention discloses a reagent set and a method for detecting avian influenza viruses and chicken parvoviruses. The reagent set for detecting or assisting in detecting the avian influenza viruses and the chicken parvoviruses comprises a primer pair named as M-P and a primer pair named as ChPV-P, the M-P comprises two single-chain DNA (deoxyribonucleic acid) shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table, and the ChPV-P comprises two single-chain DNA shown as SEQ ID No.3 and SEQ ID No.4 in the sequence table. Experimental results show that the reagent set and the method for detecting or assisting in detecting the avian influenza viruses and the chicken parvoviruses are good in specificity and accuracy and high in sensibility and can be used for specifically detecting the avian influenza viruses and the chicken parvoviruses, and the sensibility of detecting the avian influenza viruses and the chicken parvoviruses is 100fg.

The invention relates to a preparation method for obtaining blood used for preparing homologous antiserums and a spleen byproduct used for preparing transfer factors from the bodies of fur bearing animals such as foxes, raccoon dogs and minks; the furs of the foxes, the raccoon dogs and the minks are taken concentratedly in December and confirmed according to the fur-taking dates of different areas; healthy individuals are picked up 45 days before a predicated fur-taking date; the booster immunization of univalent vaccines or polyvalent vaccines of canine distemper, Canine parvovirus enteritis, adenovirus disease, corona virus laxness, parainfluenza, and pasteurellosis is carried out for 3 times by purified and condensed antigens on the foundation of carrying out immunization for once in summer; when the furs are taken, aseptic anticoagulation blood collection is carried out to a heart to improve the blood serum yield; and the spleens of the animals are collected for extracting specific or common transfer factors simultaneously. The blood can be conveniently prepared into the blood serums which resist communicable diseases, and the transfer factors can be extracted the spleen, thus providing guarantee for the healthy development of the breeding in the next year; the invention is used for curing the corresponding communicable diseases of the wild animals of the same species; and when a small amount of communicable disease cases appear in a breeding crowd, the corresponding pathogeny hyper-immune serums and transfer factors are mainly injected to a supposed healthy crowd after the pathogeny is confirmed so as to cure the affected animals in the early period of disease and control the spreading of the epidemic situations.