19 results about "Marek's disease" patented technology

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 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 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 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 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 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 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 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.

The application provides a characteristic protein of a serum type I chicken Marek's disease virus and a detection application, and belongs to the technical field of virus detection. The application provides a recombinant gene derived from the serum type I chicken Marek's disease virus, and the nucleotide sequence of the recombinant gene is shown as SEQ ID No. 1. The recombinant gene encodes MDV-1 recombinant PP38 protein, and the MDV-1 recombinant PP38 protein can be used for ELISA detection. The application also provides an ELISA detection kit constructed by using the MDV-1 recombinant PP38 protein, which can be used for clinical MDV-1 serum antibody detection, screening of SPF chickens, detection of exogenous virus antibodies of avian live vaccines or virus seeds and epidemiological investigation, so as to prevent and control the spread of MDV-1 infection.

The invention provides a porcine giant circovirus as well as a culture and separation method and application thereof, and belongs to the technical field of biotechnology and veterinary medicine. The invention provides a porcine giant circovirus gene. The nucleotide sequence of the porcine giant circovirus gene is shown as SEQ ID NO. 1. The invention also provides an application of the Marek's disease tumor cell line MSB-1 in separating and culturing the porcine giant circovirus. The porcine giant circovirus is successfully cultured on the Marek's disease tumor cell line MSB-1 for the first time, and high-titer virus liquid enough for vaccine preparation can be obtained. The porcine giant circovirus obtained by separation can be used as a totivirus inactivated vaccine, has the advantage of inducing a high-level antibody after being inactivated and added with an adjuvant to immunize a mouse, and can be subsequently prepared into a porcine circovirus type 5 inactivated vaccine. According to the invention, a key vaccine production platform is directly provided for preventing and controlling diseases such as porcine reproductive disorder caused by the porcine giant circovirus, and the blank of a porcine giant circovirus disease prevention and control technology is filled.

The invention discloses a primer pair and a kit for detecting a Marek's disease virus MEQ gene as well as a detection method and application thereof, and belongs to the technical field of biological detection. The nucleotide sequences of the forward primer and the reverse primer of the primer pair are shown as SEQ ID NO. 1 and SEQ ID NO. 2 in sequence. According to the primer pair for Marek's disease virus MEQ gene detection and the kit and the detection method based on the primer pair, based on the primer pair, the Marek's disease virus can be specifically amplified, rapid and accurate detection of the Marek's disease virus is achieved, and the primer pair has the advantages of being easy and convenient to operate, low in cost, high in sensitivity and the like; the problems that an existing detection method is high in detection cost, long in time, high in requirement for detection personnel, needs professional training, is only suitable for auxiliary diagnosis of a laboratory, cannot be popularized and applied and the like are solved, and the method has important significance on early detection of viruses.