183 results about "Canine distemper virus CDV" patented technology

Attenuated recombinant viruses containing DNA coding for a canine distemper virus antigen or measles M or N antigen, as well as methods and compositions employing the viruses, are disclosed and claimed. The recombinant viruses can be NYVAC or ALVAC recombinant viruses. The DNA can code for at least one of: canine distemper virus fusion protein, canine distemper virus hemagglutinin glycoprotein, canine distemper nucleocaspid protein, canine distemper matrix protein, measles virus nucleocaspid protein, and measles virus matrix protein. The recombinant viruses and gene products therefrom are useful for eliciting protection against canine distemper virus and / or measles virus, and, the gene products and antibodies elicited thereby are useful in assays. Additionally, DNA from the recombinants is used for probes or for generating PCR primers.

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 invention provides two mouse bone marrow hybridoma cell strains, a monoclonal antibody generated by the same and application, canine distemper virus resistant monoclonal antibodies 6E11 and 1G5 respectively secrete by the mouse bone marrow hybridoma cell strains, a prepared detecting kit and pharmaceutical composition containing the monoclonal antibodies 6E11 and 1G5, and application of the detecting kit and pharmaceutical composition, and belongs to the technical field of biology. The prepared canine distemper virus antigen detection kit containing the monoclonal antibodies 6E11 and 1G5 is high in sensitivity, and is easy and convenient to operate, quick and time-saving, or the detection rate of CDV infection is improved, application in the aspect of large-scale screening is facilitated, and the prepared canine distemper virus antigen detection kit containing the monoclonal antibodies 6E11 and 1G5 is suitable for quick detection in grassroots or in site.

Immunogenic compositions and broad-spectrum vaccines containing newly identified isolates of canine distemper virus (CDV) collected from a geographic area are provided. The newly identified isolates exhibit attributes of both European wildlife lineage CDV and one or both of Arctic and American-2 lineage CDV. Therefore, the vaccines are broadly protective against infection with European wildlife lineage CDV and either Arctic lineage CDV or American-2 lineage CDV, or both Arctic and American-2 lineage CDV.

The invention relates to a monoclonal antibody and an antibody composition for neutralizing canine distemper virus (CDV), belonging to the technical field of biology. The monoclonal antibody 2G3 disclosed by the invention has high activity for neutralizing CDV and is used for identifying epitopes differently acting on CDV with a monoclonal antibody for neutralizing CDV prepared from another screened hybridoma cell 1D7 strain. Two monoclonal antibodies are prepared into the antibody composition, the neutralizing capability on CDV of which is obviously prior to the single monoclonal antibodies 2G3 and 1D7 in the composition; after being combined, the two monoclonal antibodies can generate antiviral synergistic enhancement action; and both the range and the capability for neutralizing CDV are increased. The monoclonal antibody disclosed by the invention is used for preparing the antibody composition for treating clinical canine distemper paroxysm animal; the virus neutralizing capability is stronger; failure of the monoclonal antibody caused by CDV heteromorphosis can be avoided; and the clinical application range is wider.

The invention discloses a canine distemper attenuated vaccine strain and application thereof. In the invention, passing and cloning are performed on a separated canine distemper virulent strain to culture the canine distemper attenuated vaccine strain, and the microorganism collection number is CGMCC No.3810. The canine distemper attenuated vaccine strain of the invention can provide better protection for the canines suffering homological virulent attack, has perfect immunogenicity and can provide relatively good immune protection for the canines infected with the canine distemper virus. The attenuated vaccine strain can be prepared into single vaccine or united vaccine (live vaccine or inactivated vaccine) and can effectively prevent or cure the canine distemper. The attenuated vaccine strain of the invention has the advantages of stable transmissibility, lasting immunity, good effect, safety, reliability, long storage time and the like.

The invention discloses a ternary PCR (polymerase chain reaction) kit, which can simultaneously detect mink canine distemper virus, enteritis parvovirus and Aleutian disease virus. The ternary PCR kit comprises DNA (deoxyribonucleic acid) polymerase, dNTPs (deoxynucleotide triphosphates), primers, PCR buffer solution and double-distilled water; and the ternary PCR kit is characterized in that theprimers comprise three pairs of the primers, wherein the sequences of the first pair of the primers are 5'-GATAAAGCATGTCATTATAGTCCTAA-3' and 5'-CTTGAGCTTTCGACCTTC-3', the sequences of the second pairof the primers are 5'-AACTGGGCGGAGCCTAAA-3' and 5'-CAGAGCGAAGATAAGCAG-3', and the sequences of the third pair of the primers are 5'-GAAACCACGGTGGAGACA-3' and 5'-AAGAGTTGACCTGGAGGG-3'. By adopting themultiple PCR kit, whether the infection or the multi-infection of the mink canine distemper virus, the enteritis parvovirus and the Aleutian disease virus exists or not can be simultaneously detectedin a same reaction system, thereby having higher specificity and sensitivity, saving time and reagent and providing an effective tool for fast and early diagnosis of a mink group.

The invention relates to canine distemper live vaccine and a preparation method thereof. The preparation method takes a canine distemper virus natural attenuated strain CGMCC No.3201 with excellent immunogenicity and obtained by the inventor through field separation as a production strain to prepare the safe, effective and single-component canine distemper live vaccine. The canine distemper live vaccine effectively reduces the risk of importing other live viruses during canine distemper vaccine immunization and provides conditions for controlling the spread of fur-bearing animal diseases.

The invention discloses a mink canine distemper-parvovirus enteritis bigeminal live vaccine as well as a preparation method and application thereof. The bigeminal live vaccine comprises the effective constituents of a mink parvovirus attenuated vaccine strain MEVB-F61 and a mink canine distemper attenuated vaccine strain CDV3-CL, wherein the artificially-attenuated mink parvovirus attenuated vaccine strain MEVB-F61 is preserved in the China General Microbiological Culture Collection Center, and the preservation number of the mink parvovirus attenuated vaccine strain MEVB-F61 is CGMCC No.9560. Safety testing results show that no any adverse effect is realized when the mink is inoculated with the bigeminal live vaccine; an effect test result shows that after inoculation of the single-dose bigeminal live vaccine, the minks are prevented from attach of high toxicity for detecting canine distemper virus and parvovirus, so that minks are protected. The bigeminal live vaccine provided by the invention can simultaneously prevent two diseases of mink canine distemper and mink parvovirus enteritis, that two diseases are prevented through an injection of the bigeminal live vaccine is realized, and the bigeminal live vaccine has a wide application prospect.

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 immune colloidal gold test paper capable of simultaneously detecting canine distemper and canine parvo of foxes, raccoon dogs and minks and a method for preparing the immune colloidal gold test paper. The test paper consists of a test paper box and a test paper tape, and the test paper tape is made by sequentially bonding a hard polyvinyl chloride lining plate, a nitrocellulose membrane, a long colloidal gold combination pad, a short colloidal gold combination pad, a sample pad and an absorption pad. VP2 gene monoclonal antibody- colloidal gold biomarkers resistant to mink-source canine distemper viruses and parvoviruses are sprayed on the long and short colloidal gold combination pads respectively. Two detection lines formed by rabbit anti-mink source canine distemper virus and parvovirus antibodies and a quality control line formed by a rabbit antimouse antibody are sprayed on the nitrocellulose membrane. The test paper is applicable to quick detection of pestilences of foxes, raccoon dogs and minks, and detection of two pestilences including canine distemper and canine parvo of foxes, raccoon dogs and minks can be simultaneously completed by the same test paper, and detection is simple, convenient, quick and accurate.

The invention relates to a vaccine composition containing an immunizing amount of canine distemper virus antigens and other antigens, a preparation method and application, and relates to a preparation method and application of a kit containing an immunizing amount of canine distemper virus antigens. The prepared vaccine composition and kit can be efficiently applied to medicines for preventing and / or treating diseases related to canine distemper. The vaccine composition and kit can efficiently prevent and treat infection with popular canine distemper virus strains at the present, and can be efficiently immune to multiple pathogens.

The invention discloses a rapid quantitative detection card for a canine distemper virus antibody and a using method. The rapid quantitative detection card comprises a detection card shell and a teststrip assembled in the detection card shell. The test strip comprising a plastic base plate with pressure-sensitive adhesive. A sample pad, a marker pad, a nitrocellulose membrane and absorbent paperare sequentially pasted on the base plate. The marker pad is composed of a carrier base layer and a marker, wherein the marker is a membrane formed by spraying the carrier base layer with lanthanide fluorescent detection microspheres and lanthanide fluorescent quality control microspheres. The part, coated with a canine distemper virus H protein antigen, of the nitrocellulose membrane is a detection line. The part, coated with an anti-Chicken IgY antibody, of the nitrocellulose membrane is a quality control line. The marker is fluorescent detection microspheres marked with a canine distemper virus structural protein H protein recombinant antigen and fluorescent quality control microspheres marked with the anti-Chicken IgY antibody. By the adoption of the rapid quantitative detection card,on-site rapid quantitative determination of the canine distemper virus antibody can be achieved, and the practical value and the promotional value are higher.

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 provides a detection kit and an application thereof and belongs to the field of biological detection. The kit comprises effective doses of anti-CDV (canine distemper virus) monoclonal antibodies 1 and 2, effective doses of anti-CPIV (canine parainfluenza virus) monoclonal antibodies 3 and 4 as well as a detection reagent for detecting an antigen-antibody reaction, wherein the monoclonal antibodies 1 and 2 as well as the monoclonal antibodies 3 and 4 can be subjected to the antigen-antibody reaction with CDV and CPIV respectively at the same time. The kit can effectively detect the two viruses and antigens of the two viruses, when enzyme reaction is adopted for detection, the sensitivity of the enzyme reaction is equivalent to that of PCR (polymerase chain reaction), and when colloidal gold is adopted for detection, the sensitivity of the colloidal gold is better that that of commercially available similar products. The invention further provides the application of the kit in the aspect of non-diagnostic purpose.

The invention relates to a hybrid tumor cell strain capable of secreting high neutralizing activity CDV (canine distemper virus) monoclonal antibody. A hybrid tumor cell strain 1D7 is selected from a hybrid tumor cell bank containing 82 strains which secrete CDV monoclonal antibody, and is used for preparing mouse ascitic antibody which has neutralizing potency up to 106 to CDV. The hybrid tumor cell strain capable of secreting high neutralizing activity CDV monoclonal antibody is high in neutralizing potency and resistant to CDV strains in a wide range, and has evident treatment effect on CDattached dogs, minks or foxes in clinical treatment, and is safe without adverse side effects. The monoclonal antibody therapeutic agent prepared with the hybrid tumor cell strain has stable neutralizing potency after two years of storage, and is high in stability.

The invention discloses a canine distemper virus (CDV) sensitive cell line and an establishment method and application thereof, and belongs to the technical field of biology. According to the hundestaupe virus sensitive cell line, lentivirus four-plasmid packaging system is adopted to obtain a strain of stable cell line madin-darby canine kidney-cell isolate (MDCK-CSL) which expresses canine source signal lymphocyte activating molecules (SLAM), wherein the conservation number of strains is CGMCC No. 5881. The comparison of the propagation conditions of a CDV standard strain on the MDCK-CSL and MDCK cells indicates that after the CDV-Snyder Hill standard strain infects the MDCK-CSL cell line for 24 hours, cytopathy of the fusion, rounding contraction, obvious death and the like of cells can be observed; and the MDCK cells infected for 6 days continuously grow slowly, and the cytopathy does not occur. The CDV sensitive cell line MDCK-CSL provides a technical platform for the separation of CDV wild strains and the complete study of biological properties of the CDV wild strains, and also establishes a foundation for the prevention and control of canine distemper.

The invention provides a canine distemper virus colloidal gold immunochromatographic test strip and a preparation method thereof, relates to the technical field of colloidal gold test strip, and solves the problems of complex test procedures, time consumption, professional operation requirement and dependence on the laboratory in the existing CDV detection techniques. The test strip includes a PVC base plate, and a sample pad, a gold-labeled combined pad, a nitrocellulose membrane with a detection line and control line, and an absorption pad, which are pasted on the base plate. The gold combined pad is wrapped with a coupling marker of purified canine distemper virus monoclonal antibody N18 and colloidal gold; the detection line on the nitrate cellulose membrane is wrapped with a purified canine distemper virus purified monoclonal antibody C42; and the control line on the nitrocellulose membrane is coated with a goat-anti-mouse IgG. The test strip of the invention has the advantages of simple and rapid operation, clear and easy detection, strong specificity, high sensitivity, and no need of instruments and equipment.

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 mink viral enteritis and canine distemper binary living vaccine as well as a preparation method and application thereof. The effective components of the binary living vaccine comprise a mink viral enteritis virus (MEV) antigen and a canine distemper virus (CDV) antigen, wherein the MEV antigen is a JLM strain and has the collection number of CGMCC No. 9904, and the CDV antigen is a JTM strain and has the collection number of CGMCC No. 9905. Researches on the humoral immunity, the immunizing and virus attacking protection level and the pathologic change show that the JLM strain and the JTM strain are free from mutual immunity interference. The safety and effect of the binary living vaccine are researched further accordingly, and the research result shows that the binary living vaccine is safe to use and is not lower than the unitary living vaccine in effect.

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 relates to the field of veterinary biological products and particularly relates to a preparation method of a mink canine distemper virus live vaccine. The method comprises the following steps: inoculating a bioreactor with sensitive cells for vaccine preparation, and culturing by using a micro-carrier; after the sensitive cells are cultured by over 50% and grow into a dense single layer, inoculating the bioreactor with canine distemper virus for enrichment culture; harvesting the virus culture liquid and micro-carrier; performing freeze-thawing and removing the micro-carrier and cell debris to obtain virus liquid; and blending the virus liquid to obtain the vaccine. By improving the reaction conditions of each step and optimizing the production flow, the method provided by the invention realizes the technical effects of short production cycle, high virus titer, stable product quality, increased production efficiency and low side effect.

The invention relates to a recombination newcastle disease LaSota attenuated vaccine for expressing canine distemper virus fusion protein (F) or canine distemper virus hemagglutinin protein (H). Particularly, the recombination newcastle disease LaSota attenuated vaccine is rLa-CDVR-F or rLa-CDVR-H. The invention further discloses a method for preparing the recombination newcastle disease LaSota attenuated vaccine and application of the recombination newcastle disease LaSota attenuated vaccine in preparation of vaccines / kits.

The invention provides a method for labeling antibodies by colorful fluorescent granules and a test paper strip prepared from the antibodies. Animal viruses including, but not limited to, canine distemper viruses, canine parvoviruses, canine adenoviruses, canine coronavirus, rabies viruses, feline leukemia viruses, Marek's disease viruses, Newcastle disease viruses and the like are used as targets, and the corresponding antibodies labeled by colorful fluorescent micro-spheres are prepared by the aid of chemical covalent processes and can be applied to detecting the animal viruses. The method for labeling the antibodies by the colorful fluorescent granules and the test paper strip prepared from the antibodies have the advantages that colorful detection lines with bright developed colors can appear during detection, and accordingly qualitative results can be obtained; the contents of the animal viruses in samples further can be subsequently quantitatively obtained, accordingly, test results are clear and are high in stability, and the test paper strip can be stored at the room temperature for a long time.

The present invention relates to a method for preparing anti-canine distemper virus monoclonal antibody by adopting a bioreactor. The method is characterized by comprising the following steps: 1) carrying out serum-free suspension adaptation culture on a hybridoma cell line secreting anti-canine distemper virus monoclonal antibody; 2) culturing the hybridoma cells in a bioreactor in two stages; 3) harvesting a culture supernatant; and 4) carrying out ultra-filtration and freeze-drying to obtain the finished product. According to the method for preparing the anti-canine distemper virus monoclonal antibody by adopting the bioreactor, defects of high hybrid protein content in products, unstable quality, large batch difference and the like of the method for preparing the anti-canine distemper virus monoclonal antibody in animal bodies in the prior art are overcome, and advantages of definite culture medium components, controllable method and process, high antibody purity, convenient operation, high production efficiency and the like are provided.

The invention provides a method for producing a therapeutic monoclonal antibody of a canine distemper virus, a product thereof and a hybridoma cell. The method provided by the invention comprises the following steps: (1) carrying out amplification culture on the hybridoma cell which secretes and produces the therapeutic monoclonal antibody of the canine distemper virus in a cell culture solution; (2) then, inoculating the hybridoma cell into a cell culture solution in a bioreactor and culturing; (3) harvesting the therapeutic monoclonal antibody, produced by the hybridoma cell, of the canine distemper virus. The method provided by the invention has the advantages that the produced therapeutic monoclonal antibody of the canine distemper virus is high in purity, little in batch difference and easy in quality control, and the quality of the therapeutic monoclonal antibody can be improved remarkably.

The invention relates to the technical field of bioengineering, and particularly relates to an eukaryotic expression recombinant plasmid and a construction method thereof and a Vero cell line stably expressing the plasmid. The eukaryotic expression recombinant plasmid has the nucleotide sequence shown in SEQ ID NO:1. The Vero cell line stably expressing the plasmid can be used for high-efficiency separation of canine distemper virus virulent strains and the research of the pathogenic mechanism of canine distemper virus.

The invention discloses a Taqman probe fluorescent quantitative PCR detection kit for identifying a canine distemper virus wild strain and a vaccine strain as well as application thereof. The kit comprises a universal primer for amplifying a canine distemper virus wild strain and vaccine strain H gene conservation area as well as a specific probe for identifying a canine distemper virus wild strain or a vaccine strain. The experiment proves that the Taqman probe fluorescent quantitative PCR detection kit is used for detecting and identifying the CDV wild strain and the vaccine strain, the sensitivity can reach to 1 copy; 103 to 102 copies with the virus quantity as low as 1 TCID50 / mL and higher than that of the conventional method can be detected; and through detection on five kinds of other pathogen nucleic acid, the copies are negative. Therefore, the kit for identifying the CDV wild strain and the vaccine strain has the characteristics of high specificity, high sensitivity and highstability, and can rapidly identify canine distemper virus wild strain and vaccine strain infection.

The invention provides a Canine-SLAM / Vero cell line and a constructing method, wherein the constructing method comprises the following steps of: collecting canine distemper positive canine anticoagulated blood, separating peripheral blood lymphocytes by applying a canine lymphocyte separation liquid, extracting total RNA (Ribonucleic Acid), and obtaining a canine SLAM gene by uisng a PCR (Polymerase Chain Reaction) method; constructing pCAGGS-Neo / SLAM plasmids; transfecting a constructed pCAGGS-Neo / SLAM expression cassette into a Vero cell, and screening the cell line for stably expressing an SLAM by adopting G418; and identifying and detecting the expression condition of the SLAM on the surface of a cell membrane by utilizing an indirect immunofluorescent method to obtain the cell line. The invention not only can be used for the separation of canine distemper viruses and the titration of the viruses, but also markedly shortens the time for separating the viruses, improves the yield of the viruses, and forcefully promotes researches on the molecular characteristics, such as phenotypes, genotypes and the like of the canine distemper viruses.

The invention provides a primer set, a reagent kit and a method for detecting canine distemper viruses (CDV), canine parvoviruses (CPV) and canine coronaviruses (CCV), and belongs to the technical field of virus detection. The primer set comprises primers with nucleotide sequences shown as SEQ ID No.1-6. The primers correspond to three pairs of upstream and downstream primer pairs for the canine distemper viruses (CDV), the canine parvoviruses (CPV) and the canine coronaviruses (CCV). The reagent kit comprises the primer set. The method is based on the primer set and the reagent kit and is used for detecting the canine distemper viruses, the canine parvoviruses and the canine coronaviruses. The primer set, the reagent kit and the method have the advantages that the canine distemper viruses, the canine parvoviruses and the canine coronaviruses can be detected by the aid of the primer set, the reagent kit and / or the method, the primer set, the reagent kit and the method are good in specificity and accuracy and high in sensitivity and efficiency and are simple and speedy, reliable technical means can be provided to clinical detection, and the like.