37 results about "Marek's disease" patented technology

The invention belongs to the field of veterinary biotechnology detection, and particularly relates to a hybridoma cell strain and a monoclonal antibody for detecting or identifying MDV-1 and application of the hybridoma cell strain and the monoclonal antibody. The hybridoma cell strain MDV-1Mab-pp38-E2 is obtained through screening by taking pp38 protein of MDV-1 as immunogen, the preservation number is CCTCC NO: C202501, the monoclonal antibody generated by the hybridoma cell strain can recognize I-type chicken Marek's disease virus strains with different serum, and the monoclonal antibody can be used for identifying the type I chicken Marek's disease virus strains with different serum types. The kit has good specificity and sensitivity and can be used for exogenous virus detection of the serum type I chicken Marek's disease virus in poultry virus live vaccines without cross reaction with serum type II chicken Marek's disease virus, serum type III chicken Marek's disease virus, chicken egg drop syndrome virus (EDSV) and other common poultry disease viruses, and the kit has good specificity and sensitivity and can be applied to exogenous virus detection of the serum type I chicken Marek's disease virus in poultry virus live vaccines. The kit can also be used for clinical identification, virus content determination and epidemiological investigation of serotype I chicken Marek's disease virus.

The invention provides a characteristic protein of serotype I chicken Marek's disease virus and detection application, and belongs to the technical field of virus detection. The invention provides a recombinant gene derived from serum I type chicken Marek's disease virus. The nucleotide sequence of the recombinant gene is shown as SEQ ID No. 1. The recombinant gene disclosed by the invention is used for coding the MDV-1 recombinant PP38 protein, and the MDV-1 recombinant PP38 protein can be applied to enzyme-linked immuno sorbent assay (ELISA) detection. The invention also provides an ELISA detection kit constructed by using the MDV-1 recombinant PP38 protein, and the ELISA detection kit can be used for clinical MDV-1 serum antibody detection, SPF chicken screening, exogenous virus antibody detection of poultry live vaccines or virus seeds, epidemiological investigation and the like, so as to prevent and control transmission of MDV-1 infection.

The invention relates to detection of vaccine immune effects, in particular to a Marek's disease virus specificity IFN-gamma sandwich ELISA method and application thereof. The invention provides a Marek's disease virus specific T cell immune stimulator, and the amino acid sequence of the Marek's disease virus specific T cell immune stimulator is shown as SEQ ID NO: 1 or SEQ ID NO: 3. The invention further provides a method for detecting the cellular immune level after Marek's disease vaccine immunization. The Marek's disease virus specific T cell immunostimulant can effectively stimulate cell-mediated immune response, initiate and activate MDV specific T cells, so that the Marek's disease virus specific T cells generate Marek's disease virus specific IFN-gamma, and the Marek's disease virus specific T cell immunostimulant has a good application prospect in Marek's disease vaccine immune level detection.

The invention discloses a composition, a kit and a detection method for detecting Marek's disease virus, and belongs to the technical field of molecular biology diagnos.The nucleic acid composition comprises an RPA primer pair and sgRNA, the RPA primer pair is composed of an upstream primer and a downstream primer, the nucleotide sequence of the upstream primer is as shown in SEQ ID NO.1, the nucleotide sequence of the downstream primer is as shown in SEQ ID NO.2, and the sgRNA pair is composed of sgRNA and sgRNA. The RPA primer pair contains a guide sequence for specifically recognizing a target amplified by the RPA primer pair; the primer group has the advantages of high specificity and high sensitivity, and the kit designed based on the nucleic acid composition has the advantages of simplicity in operation, high detection speed, result visualization and the like.

The invention relates to the fields of molecular biology and virology, and provides a method for preparing a specific MHC haplotype-derived cell-binding virus. The method comprises the following steps: firstly, obtaining chick embryos of different haplotypes, separating CEF cells, transiently expressing exogenous fluorescent protein through electrotransfection, then carrying out virus inoculation, carrying out flow sorting to obtain a fluorescent positive cell population carrying MDV, and carrying out quantitative analysis on the cell population to obtain the specific MHC haplotype chicken Marek's Disease Virus (MDV), thereby obtaining the specific MHC haplotype chicken Marek's Disease Virus Marek's Disease Virus Marek's Disease Virus Marek's Disease Virus. And carrying out in-vitro propagation to obtain the purified MDV with a specific haplotype source. The purified virus can provide a test material for related research of haplotype on virus infection, and can be widely applied to in-vitro purification of different cell binding viruses and related research of specific MHC molecule presenting virus peptides. The method disclosed by the invention is beneficial to carrying out related research aiming at the host immune response of the virus and analyzing the hereditary basis of haplotype to MDV differential resistance, and is expected to be applied to preparation of vaccine virus.

The invention provides a molecular diagnosis method for avian leukosis virus and Marek's disease virus mixed infection. The molecular diagnosis method comprises the following steps: (S01) collecting diseased material tissues; (S02) carrying out nucleic acid extraction on the sick material tissue of the sick chicken; (S03) detecting the nucleic acid by using specific primers of avian leukosis and Marek's disease; (S04) preparing a PCR (Polymerase Chain Reaction) detection system, and respectively adding ALV-env gene amplification primers and MDV-meq gene amplification primers for gene amplification; (S05) carrying out 1% agarose gel electrophoresis detection on the amplification product in the step (4), and carrying out sequence determination after determining a target band; (S06) carrying out BLAST comparison on the sequences obtained by sequencing, carrying out gene sequence analysis and drawing an evolutionary tree; and (S07) determining the virus type according to the homology and the evolutionary branches, so as to realize rapid diagnosis of ALV and MDV mixed infection, strain characteristic analysis and exclusion of other similar epidemic diseases, and provide technical support for accurate prevention and control of AL and MD in the poultry industry, provenance purification and epidemiological investigation.

The present application belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition, a preparation method and application thereof. Specifically, based on the theory of traditional Chinese veterinary medicine of'strengthening the body resistance and eliminating pathogenic factors, and treating the root and branch', aiming at the pathogenesis of 'deficiency in the root and excess in the branch' of low immunity in animals, eight kinds of medicinal materials are scientifically screened to form a traditional Chinese medicine compound with the effects of tonifying qi and nourishing blood, invigorating the spleen and kidney, dredging collaterals and eliminating dampness, and clearing heat and detoxifying. Through the verification of a cyclophosphamide-induced low immunity mouse model, the traditional Chinese medicine composition can significantly improve the mental state of the mice, restore the body weight and thymus / spleen index, repair the pathological damage of the spleen tissue, and adjust the inflammatory factor level in the serum and spleen tissue. In addition, in a Marek's disease virus infection model, the traditional Chinese medicine composition combined with the Marek's disease virus vaccine can significantly reduce the viral load in the peripheral blood, liver and spleen, and reduce the liver enlargement, hemorrhage and microscopic structure damage, showing a good immune synergistic effect.

The present application belongs to the field of veterinary biotechnology detection, and particularly relates to a hybridoma cell strain for detecting or identifying MDV-1, a monoclonal antibody and application thereof. The present application takes pp38 protein of MDV-1 as an immunogen to screen a hybridoma cell strain MDV-1 Mab-pp38-E2 with a preservation number of CCTCC NO: C202501, and the monoclonal antibody generated by the hybridoma cell strain can recognize different serum type I chicken Marek's disease virus strains, does not have cross reactions with serum type II chicken Marek's disease virus, serum type III chicken Marek's disease virus, chicken egg drop syndrome virus (EDSV) and other common avian disease viruses, has good specificity and sensitivity, can be used for exogenous virus detection of serum type I chicken Marek's disease virus in avian virus live vaccines, and can also be used for clinical identification, virus content determination and epidemiological investigation of serum type I chicken Marek's disease virus.

The present invention provides an effective vaccine for Marek's disease, which may be prepared using a recombinant Marek's Disease Virus (MDV), strain CVI988, having been transformed with a foreign DNA construct that includes a long terminal repeat sequence of a reticuloendotheliosis virus. This safe viral agent elicits a highly protective immune response in a chicken against virulent MDV challenge without causing a significant degree of pathogenicity. Suitable formulations of the vaccine for use in chickens include an effective immunization dosage of this novel viral agent, along with a pharmaceutically acceptable carrier or diluent.

The invention belongs to the technical field of biological vaccines, and particularly relates to a chicken Marek's disease virus miR-M6-5p gene deleted strain and application thereof. The invention provides a chicken Marek's disease virus miR-M6-5p gene deleted strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.46202. The invention further provides a vaccine containing the chicken Marek's disease virus miR-M6-5p gene deletion strain and a preparation method of the vaccine. The replication level of the gene deletion strain on cells is remarkably increased, and the incidence of tumors caused by infection of chickens is reduced, so that the gene deletion strain can be used for research and development of chicken Marek's disease vaccines and has a good application prospect in prevention of chicken Marek's disease.

The invention belongs to the technical field of biology, and particularly relates to an anti-Marek's disease virus US3 protein monoclonal antibody and application thereof. The invention provides an anti-Marek's disease virus US3 protein monoclonal antibody which is secreted by a hybridoma cell with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.46324. The antibody is strong in specificity and high in sensitivity, can be used for directly detecting the MDV US3 protein by an immunofluorescence method, realizes rapid diagnosis of MDV infection, and provides technical support for further researching the biological function of the MDV US3 protein and further revealing an MDV pathogenic mechanism.

The present invention provides a recombinant Marek's disease virus that co-expresses the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene and its application, belonging to the field of genetically engineered vaccines. The present invention utilizes recombinant cloning technology to insert an expression framework comprising the Newcastle disease virus gene type VII strain F gene and the infectious bursal disease virus new variant VP2 gene into the serotype 1 chicken Marek's disease virus CVI988 strain genome between nucleotides 105561-105563, constructing a recombinant cosmid in which the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene expression framework are inserted into the chicken Marek's disease virus genome, and the recombinant virus C20103 strain that co-expresses the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene is rescued by the recombinant Marek's disease virus. The vaccine prepared by the recombinant chicken Marek's disease virus provided by the present invention can simultaneously prevent Newcastle disease, infectious bursal disease and chicken Marek's disease, and has good market application prospects.

The invention discloses a recombinant Marek's disease virus for expressing a serum 4 type fowl adenovirus Fiber2 gene as well as a construction method and application of the recombinant Marek's disease virus, and belongs to the technical field of biology. The recombinant Marek's disease virus is obtained by inserting a gene segment SV40-Fiber2 containing an SV40 promoter sequence, an FAdV-4 Fiber2 gene and a terminator sequence into a UL41 gene of a Marek's disease virus Meq gene deleted vaccine strain. A challenge protection test result shows that after chicken is immunized by the recombinant virus rMDV-4 Fiber2, good protection can be provided for FAdV-4 and MDV challenge, and 80% protection and 100% protection are respectively provided for death caused by FAdV-4 virulent strain and MDV virulent attack. The construction of the recombinant virus rMDV-Fiber2 provides a technical means for the development of HHS recombinant MDV live vector vaccines, and is of great significance to the prevention and control of HHS.

The present invention relates recombinant viruses and the uses thereof. More particularly, the invention relates to novel recombinant Marek's disease viruses encoding polypeptide(s) of interest, and their use to express or deliver such polypeptides to animals, particularly poultry. The invention is particularly suited to vaccinate poultry against avian pathogens.

The application provides a Marek's disease virus meq gene edited deletion vaccine candidate strain SQ01Delta meq and construction and application. The SQ01Delta meq vaccine candidate strain has a preservation number of CGMCC NO.45864. The parent strain of the SQ01Delta meq vaccine candidate strain is MDV super strong Chinese mutant HNSQ01. After in vitro passage of the parent strain HNSQ01, the CRISPR / Cas9 gene editing technology is used to delete a 925bp fragment on each of the two meq alleles in the viral genome. Through PCR identification, gene cloning sequencing, RT-qPCR analysis, IFA identification and SPF chicken challenge experiment, it is ensured that the SQ01Delta meq strain completely loses the pathogenicity and tumorigenicity to the host chicken, and has no immunosuppressive property, and has good biosafety. Animal immune challenge protection evaluation experiment shows that the SQ01Delta meq has good immune protection, and the immune protection index of the parent strain reaches 80.0%. Therefore, the SQ01Delta meq strain constructed by the application can not only be used as a new MD gene deletion vaccine candidate strain, but also be used for the development and utilization of subsequent MD new type gene engineering vaccine.

The application provides an amplification primer and method for a whole genome of chicken Marek's disease virus. The detection primer set of the chicken Marek's disease virus comprises 206 pairs of amplification primers, and specific sequences are shown as SEQ ID NO. 1-412. The detection primer set provided by the application can realize relatively uniform coverage of the chicken Marek's disease virus, and in nanopore sequencing, 100% area coverage of the genome is realized. The sequencing fragment preparation method for the whole genome of the chicken Marek's disease virus provided by the application is simple and easy to operate, has good amplification effect, and nanopore sequencing has the advantage of real-time analysis during sequencing, so that the detection time can be greatly shortened, and the infection of the chicken Marek's disease virus can be differentially diagnosed in 15h at most, meanwhile, the whole genome sequence thereof is obtained, and the obtained genome sequence can provide a scientific basis for virus tracing, pathogen variation tracking, new strain identification and early warning and the like.

The invention relates to a diluent of a Marek's disease live vaccine and a preparation method of the diluent, and aims to improve the immune effect of the vaccine and relieve the immune stress reaction after inoculation. The diluent takes water for injection as a solvent and comprises the following components: cane sugar, L-sodium glutamate, dipotassium phosphate, monopotassium phosphate, sodium chloride, beef extract, phenol red, water-soluble yeast glucan, meglumine and the like. The mass stability and the biological safety of the diluent are ensured by accurately controlling the proportion of each component and strict preparation processes including raw material weighing, dissolving and mixing, pH adjusting, filtering and purifying and storing. Experimental results show that the vaccine diluted according to the optimized formula can effectively improve immune protection, meanwhile, the influence on the weight and immune organs of test chickens is reduced, the obvious anti-stress characteristic is shown, and the vaccine has a good application prospect.

The invention relates to the technical field of gene engineering, and discloses application of an inhibitor of LRP1 protein or a coding gene thereof in preparation of a preparation for resisting MDV infection. Specifically, the invention discloses application of the LRP1 protein or the coding gene thereof in regulation and control of Marek's disease virus (MDV) in-vitro infection of chicken embryo fibroblasts (CEF). The invention finds that the MDV infection rate can be remarkably reduced by knocking down the LRP1 gene in CEF cells, and further provides application of the LRP1 protein or the coding gene thereof or a biological material containing the coding gene thereof in resisting MDV infection. According to the invention, the effect of the LRP1 gene in the in-vitro infection of the CEF by the MDV is defined, and a foundation is laid for further research on the antiviral mechanism of the LRP1 gene and application of the LRP1 gene in disease-resistant breeding.

The invention discloses a Marek's low virulent strain, the whole genome sequence of the Marek's low virulent strain is characterized in that a nucleotide sequence as shown in SEQ ID NO.1 is deleted behind the 123439th basic group of a Marek's virus gene sequence DQ530348.1, and a long terminal repetitive sequence of avian reticuloendotheliosis virus as shown in SEQ ID NO.2 is inserted behind the 123439th basic group. The invention also discloses a construction method of the Marek's low virulent strain rescue system and application of the Marek's low virulent strain in preparation of vaccines for preventing or treating Marek's disease. The Marek's attenuated strain disclosed by the invention can quickly generate antibodies, has the detoxification capability obviously weaker than that of a vaccine virus CVI988 / Rispens strain, is a high-quality vaccine strain for developing Marek's disease vaccines, and has a very good application prospect.

The invention provides a recombinant Marek's disease virus strain for co-expressing a Newcastle disease virus F gene and an H9 subtype avian influenza virus HA gene and application, and belongs to the field of genetic engineering vaccines. According to the invention, by using a recombinant cloning technology, an expression framework containing a Newcastle disease virus gene VII type strain F gene and an H9 subtype avian influenza virus HA gene is inserted into a spacer region between Marek's disease virus genome UL45 and UL46 genes; according to the construction method, the recombinant cosmid is obtained by inserting a Newcastle disease virus F gene and an H9 subtype avian influenza virus HA gene expression framework between Marek's disease virus genome UL45 and UL46 genes, and the recombinant virus for co-expressing the Newcastle disease virus F gene and the H9 subtype avian influenza virus HA gene is obtained by rescue of the recombinant cosmid. The vaccine prepared from the recombinant Marek's disease virus strain provided by the invention can prevent Newcastle disease, H9 subtype avian influenza and Marek's disease at the same time, and has a good market application prospect.

The invention discloses a recombinant Marek's virus vector vaccine strain for expressing H9N2 subtype avian influenza virus HA protein and application of the recombinant Marek's virus vector vaccine strain, and belongs to the technical field of veterinary biological products. The amino acid sequence of the HA protein expressed by the recombinant Marek's virus vector vaccine strain is as shown in SEQ ID NO. 1. The amino acid sequence of the HA protein is obtained by modifying the 84th amino acid of the amino acid sequence shown as SEQ ID NO.2 into tryptophan from serine, eliminating the 82nd glycosylation site of the HA protein, replacing the signal peptide of the HA gene with the Ig lambda signal peptide of chicken, and adding GGGGS linker and one tetanus toxin T cell epitope to the carboxyl terminal of the HA protein. The vaccine strain is good in in-vitro proliferation characteristic, stable in genome heredity and capable of stably expressing HA protein, no side reaction or pathogenic effect exists after SPF chicken is immunized, 100% protection against H9N2 strain attack can be provided 21 days after immunization, and the problem that the immune protection blank period of other vaccines such as H9N2 inactivated vaccines is long is solved.

The present invention discloses novel recombinant multivalent non-pathogenic Marek's Disease virus constructs that encode and express both Infectious Laryngotracheitis Virus protein antigens and an Infectious Bursal Disease virus protein antigen, and methods of their use in poultry vaccines.

The present invention relates to the field of animal health. Particularly, the present invention relates to recombinant serotype 1 Marek's Disease Virus (rMDV-1) comprising an expression cassette in the genome thereof, wherein the expression cassette comprises a coding sequence of at least one antigenic protein of Infectious laryngotracheitis virus (ILTV) operably linked to a corresponding native ILTV promoter. Further, the present invention provides an immunogenic composition comprising the rMDV-1 of the present invention and the use of the immunogenic composition for preventing and / or treating diseases in an animal.

The present invention relates to the veterinary field in general and specifically to the development of vectored vaccines using the Gallid alphaherpesvirus 3 (GaHV-3) or Marek's disease serotype 2 (MDV-2) vector, which contains protective antigens against different avian pathogens such as Newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), infectious bursal disease virus (IBDV) or Gumboro disease, avian influenza virus (AIV), and infectious bronchitis virus (IBV).

The present invention relates to recombinant Marek's disease viruses containing, inserted into at least one insertion site, at least one recombinant nucleotide sequence encoding an antigen operably linked to a promoter and an insulator, their manufacture, compositions comprising the same, and the uses thereof.

The invention discloses a recombinant Marek's virus strain for co-expressing an H9 subtype AIV HA gene and an IBV S gene as well as a construction method and application of the recombinant Marek's virus strain, and belongs to the technical field of medicine or veterinary medicine. The recombinant Marek's virus is obtained by co-transfecting CEF cells with recombinant coplasmids containing HA gene and S gene expression cassettes and recombinant coplasmids containing MDV genome DNA fragments and then carrying out virus rescue. Specifically, by using a recombinant cloning technology, an expression cassette containing an H9 subtype AIV HA gene and an IBV S gene is inserted between 105561-105563 nucleotides of a serum type 1 MDV attenuated live vaccine CVI988 strain, so as to construct and obtain a recombinant clone in which the HA gene and the S gene are inserted in an MDV genome, and the recombinant clone rescues to obtain a recombinant MDV vaccine strain C20301 for co-expressing the AIV HA gene and the IBV S gene. The invention also relates to application of the recombinant MDV vaccine strain in preparation of vaccines for simultaneously preventing H9 subtype avian influenza, infectious bronchitis and Marek's disease.

The application discloses a PCR primer and probe for identifying serum type I Marek's vaccine virus and wild virus, wherein the primer comprises an upstream primer with a sequence as shown in SEQ ID No:1 and a downstream primer with a sequence as shown in SEQ ID No:2; and the probe comprises a wild virus-specific probe with a sequence as shown in SEQ ID No:3 and a vaccine virus-specific probe with a sequence as shown in SEQ ID No:4. Through the primer and the probe, the serum type I Marek's vaccine virus and the wild virus can be efficiently, conveniently and quickly identified, so that early warning, accurate prevention and control and purification of Marek's disease are realized.

The present invention relates to the field of animal health. Particularly, the present invention relates to recombinant serotype 1 Marek's Disease Virus (rMDV1) comprising an expression cassette in the genome thereof, wherein the expression cassette comprises a coding sequence of at least one antigenic protein of Newcastle disease virus (NDV). Further, the present invention provides an immunogenic composition comprising the rMDV1 of the present invention and the use of the immunogenic composition for preventing and / or treating diseases in an animal.

The present invention discloses novel recombinant multivalent non-pathogenic Marek's Disease virus constructs that encode and express foreign antigens from three or more avian viruses, along with methods of the use of the multivalent poultry virus vaccines.

The present invention relates to the field of animal health. Particularly, the present invention relates to a recombinant Marek's Disease Virus (MDV) comprising at least one exogenous nucleotide sequence in the genome thereof, wherein the at least one exogenous nucleotide sequence is located at a position between UL55 and Lorf10, or a position between UL2 and UL3. Further, the present invention provides an immunogenic composition comprising the recombinant MDV of the present invention and the use of the immunogenic composition for preventing and / or treating diseases in an animal.