17 results about "Goose parvovirus" patented technology

The invention provides a novel goose parvovirus attenuated strain as well as a construction method and application thereof, and belongs to the field of viral vaccines. Two artificial attenuating sites capable of weakening the virulence of the novel goose parvovirus are determined through research, the live vaccine prepared by taking the obtained NGPV-NMG50 attenuated strain as an immunogen does not need to add adjuvants, the defeathering protection rate of a duck group can reach 100%, the minimum lethal dose of the virus is determined by carrying out multiple dilution on an NMG21 wild strain, and the vaccine can be used for preparing the novel goose parvovirus by taking the NGPV-NMG50 attenuated strain as an immunogen. The optimal immune dose of the NGPV-NMG50 attenuated strain is screened, so that the optimal immune dose result of the NGPV-NMG50 attenuated strain in clinical use is obtained, and an immune barrier for the novel goose parvovirus is effectively established for the ducklings.

The invention discloses a conservative B cell epitope peptide of goose parvovirus VP3 protein, a nucleic acid molecule, a recombinant vector and application thereof. Conservative B cell epitope EW12 is inserted into F18 Escherichia coli Fod fimbriae, inert carrier bacteria S9H are introduced, recombinant bacteria capable of functionally exhibiting and expressing the EW12 conservative B cell epitope on the surface of the bacteria are obtained, the recombinant bacteria can specifically generate specific binding reaction with gosling plague egg yolk antibody and gosling plague vaccine immune serum, and the gosling plague egg yolk antibody and the gosling plague vaccine immune serum can be specifically combined with the gosling plague egg yolk antibody and the gosling plague vaccine immune serum. The method has the advantages that macroscopic agglutination particles are generated, cross agglutination reaction with other pathogen positive serum and healthy goose serum is avoided, the method is good in sensitivity, and the specific antibody can be detected in the 5th day after immunization at the earliest. A plate agglutination test directly mediated by the goose parvovirus VP3 protein conservative B cell epitope peptide EW12 has the advantages of specificity, sensitivity, convenience in operation and suitability for on-site large-scale rapid detection, and is expected to provide a new technical means for diagnosis, prevention and control and purification of GPV infection.

The present invention provides a novel attenuated strain of goose parvovirus and its construction method and application, belonging to the field of virus vaccines. The present invention has determined two artificial attenuation sites that weaken the virulence of the novel goose parvovirus through research. The NGPV-NMG50 attenuated strain obtained in this application is used as a live vaccine prepared as an immunogen, and the feather loss protection rate of ducks can reach 100% without the addition of adjuvants. The minimum lethal dose of the virus is determined by diluting the NMG21 wild strain in multiples, and then the optimal immune dose of the NGPV-NMG50 attenuated strain is screened, and the optimal immune dose results of the NGPV-NMG50 attenuated strain in clinical use are obtained, so that ducklings can effectively establish an immune barrier against the novel goose parvovirus.

The present invention provides a novel goose parvovirus, an inactivated vaccine and an application thereof, and belongs to the technical field of veterinary biological products. The present invention isolates and obtains a novel goose parvovirus, and poultry will develop neurological symptoms 2-3 days after being infected by the strain. The present invention inactivates the novel goose parvovirus, and the prepared novel goose parvovirus inactivated vaccine has good safety, can provide effective immune protection for ducks, and has a good prospect for commercial development. When preparing the novel goose parvovirus inactivated vaccine, the present invention first inoculates the novel goose parvovirus into DEF cells in the S phase to obtain a virus liquid, and on this basis, selects a suitable formula ratio to prepare an oil phase and an aqueous phase, and performs a fully emulsified preparation method, which improves the safety and stability of the prepared vaccine while improving the efficiency of the inactivated vaccine preparation.

The invention relates to a primer and a locked nucleic acid probe set for real-time fluorescent quantitative PCR detection of two goose parvovirus virulent and attenuated strains, the goose parvovirus virulent and attenuated strains comprise MDGPV virulent and attenuated viruses and SBDSV virulent and attenuated viruses, and the sequences of the primer and the locked nucleic acid probe set are respectively shown as SEQ ID NO.1-8. The LNA-TaqMan real-time fluorescent quantitative PCR detection method established by utilizing the primers and the LNA-TaqMan probe group can be used for simultaneously detecting strong and weak goose parvovirus strains (strong and weak muscovy duck gosling plague virus and / or strong and weak duck short beak and dwarf syndrome virus), identifying, diagnosing and accurately quantifying, and is high in sensitivity, strong in specificity and good in repeatability.

The application discloses a novel goose parvovirus VP1 protein truncated body gene, a recombinant duck plague virus and application. The novel goose parvovirus VP1 protein truncated body gene has a nucleotide sequence shown as SEQ ID No. 1. The recombinant duck plague virus has the novel goose parvovirus VP1 protein truncated body gene inserted into the genome of the recombinant duck plague virus. The application obtains a novel goose parvovirus VP1 protein truncated body gene, inserts the truncated body into the genome of the recombinant duck plague virus, and obtains the recombinant duck plague virus, and the rsVP1-Cre immune group shortens the NGPV discharge in the duck body, and the duck body immune rsVP1-Cre is helpful to the NGPV discharge. It is shown that the recombinant duck plague virus with the VP1 protein truncated body gene inserted can be used for preparation of a novel goose parvovirus-duck plague virus double live vector vaccine.

This invention discloses a fusion protein expressing novel duck reovirus σC protein and duck-derived goose parvovirus VP1 protein, a recombinant duck viral enteritis virus, a vaccine, its construction method, and its applications, belonging to the field of recombinant virus technology. The nucleotide sequence of the fusion protein is shown in positions 300-3568 of SEQ ID NO:1. The fusion protein of this invention can simultaneously express protective antigen proteins of novel duck reovirus (NDRV) and duck-derived goose parvovirus (DGPV). Using CRISPR / Cas9 gene editing technology, the fusion protein is recombined into the US3 gene of duck viral enteritis virus. The resulting recombinant duck viral enteritis virus not only provides protection against virulent duck viral enteritis virus challenge but also provides good protection against NDRV and DGPV, achieving a multi-protection effect with a single injection.

The application discloses a recombinant novel goose parvovirus infectious clone plasmid and application, the infectious clone plasmid includes the full-length genome of novel goose parvovirus, and a synonymous mutation marker is introduced at the cysteine codon position in the non-structural protein coding region, under the condition that the amino acid sequence is not changed and the offspring virus replication and infection ability are not influenced, a stable single nucleotide genetic marker is provided for the novel goose parvovirus.The infectious clone plasmid can be used for rescuing the recombinant virus carrying a specific synonymous mutation marker, the obtained recombinant virus has good duck embryo adaptability and cell infection ability, and is suitable for application scenes such as cell infection and immunofluorescence detection.The application provides an important technical tool for novel goose parvovirus genetic marker tracking, pathogenic mechanism research, genetic variation analysis and vaccine research and development.

The invention discloses a primer and a probe for real-time RPA (recombinase polymerase amplification) detection of goose parvovirus and a detection method. The primer comprises an upstream primer G-F1 and a downstream primer G-R1, the sequences of the upstream primer G-F1 and the downstream primer G-R1 are as shown in SEQ ID No.1-2, the probe is an Exo probe, and the sequence of the Exo probe is as shown in SEQ ID No.3. The detection limit of the detection method established by using the primer and the probe is as low as 3.0 * 10 < 1 > copy number / mu L, cross reaction with other waterfowl viruses is avoided, the clinical coincidence rate reaches 98.1%, detection is completed within 30 min in the whole process, a thermal cycler is not needed, and the method is suitable for field application. The matched kit comprises a primer pair, a probe and an RPA basic reagent, is simple and convenient to operate and controllable in cost, can be used for rapid diagnosis and epidemiological monitoring of GPV classical strains and variants (MDGPV and SBDSV), and has important application value.

The invention relates to an application of fucoidin (FUC) in preparation of a preparation for resisting goose parvovirus (GPV) infection, primary muscovy duck embryo small intestine epithelial cells (PMDIECs) and GPV PT isolates are used as research objects, and through the technologies of cell activity detection, immunoblotting, absolute fluorescence quantification qPCR, half tissue cell infection dose, immunofluorescence analysis and the like, the preparation of the preparation for resisting goose parvovirus (GPV) infection is prepared. The system carries out FUC cytotoxicity determination, in-vitro anti-GPV infection evaluation and action mechanism exploration. Results show that no obvious cytotoxicity exists when the FUC with the concentration of 6 mg / mL acts on PMDIECs for 72 h, the FUC can inhibit GPV infection in a dose-dependent mode, GPVVP3 protein expression in cells, the virus genome copy number, the culture supernatant virus titer and the like are remarkably reduced, it is confirmed for the first time that the FUC has the anti-GPV activity in vitro, and an experimental basis is provided for research and development of veterinary preparations for preventing and controlling GPV infection.

The invention discloses an antigen gene and application of polypeptide thereof in preventing goose parvovirus disease. The antigen gene and the polypeptide provided by the invention can be used as a neutralizing epitope protein vaccine and a neutralizing epitope gene vaccine; the neutralizing epitope protein vaccine is a protein composed of 305 amino acid residues; a gene for coding the antigen polypeptide contains 918 nucleotides. The antigen gene and the polypeptide, namely the neutralizing epitope protein vaccine and the neutralizing epitope gene vaccine, provided by the invention have the advantages of strong antigenicity, capability of inducing an organism to generate a higher neutralizing antibody, good prevention effect, no pathogen transmission and the like, and a new situation is opened up in the GP prevention history.

The invention discloses a recombinant serum type 4 fowl adenovirus for expressing goose parvovirus virus VP3 protein as well as a construction method and application of the recombinant serum type 4 fowl adenovirus. According to the recombinant serum type 4 fowl adenovirus, a coding gene for expressing goose parvovirus GPVSJ20190420 strain VP3 protein is used for replacing 253-site nucleotide to 1440-site nucleotide of a Fiber-2 gene of FAdV-4. The recombinant FAdV-4 virus FAdV4-VP3 (GPV) for expressing goose parvovirus GPVSJ20190420 strain VP3 protein is successfully constructed by taking FAdV-4 as a virus vector, targeting Fiber-2 through a CRISPR / Cas9 technology and a Cre-loxP recombinant system and utilizing a double-fluorescence system, technical support and vaccine candidates are provided for effective joint prevention and joint control of GPV and FAdV-4, and the recombinant FAdV4-VP3 (GPV) has good market application value.

The invention relates to application of fucoidan polysaccharide sulfate in preparation of an inflammation protective agent and an intestinal flora regulator for young muscovy ducks against goose parvovirus infection. The invention provides application of fucoidan polysaccharide sulfate in preparation of a young muscovy duck anti-goose parvovirus infection preparation. The application specifically comprises an anti-inflammatory protective agent for young muscovy ducks against goose parvovirus infection and a flora regulator for repairing and regulating intestinal flora. The protection effect and mechanism of the fucoidan polysaccharide sulfate on the GPV infected young muscovy ducks are explored, and in-vitro and in-vivo experiments are respectively carried out. Results show that medium and high doses of fucoidan polysaccharide sulfate can significantly reduce the content of pro-inflammatory factors in serum and target organs, recover the level of anti-inflammatory factors and relieve tissue inflammatory damage. Microflora analysis results show that medium and high doses of fucoidan polysaccharide sulfate can reverse intestinal flora diversity decline caused by GPV infection, regulate abundance of dominant phylum such as phylum firmicalis and phylum bacteroides, and enrich beneficial bacteria such as Ackermann muciniphila.

The invention discloses a fusion protein for expressing a novel duck reovirus sigma C protein and a duck-origin goose parvovirus VP1 protein, a recombinant duck virus enteritis virus, a vaccine, a construction method and application, and belongs to the technical field of recombinant viruses. The nucleotide sequence of the fusion protein disclosed by the invention is shown as the 300th to 3568th sites of SEQ ID NO: 1. The fusion protein can simultaneously express protective antigen proteins of a novel duck reovirus (NDRV) and a duck-origin goose parvovirus (DGPV), the fusion protein is recombined into a US3 gene of the duck viral enteritis virus by utilizing a CRISPR / Cas9 gene editing technology, and the obtained recombinant duck viral enteritis virus can provide protection for virulent attack of the duck viral enteritis virus, and can also provide a protective effect for the duck viral enteritis virus. And a good protection effect on the NDRV and the DGPV can also be achieved, and the one-needle multi-prevention effect is achieved.

The invention provides a novel goose parvovirus, an inactivated vaccine and application thereof, and belongs to the technical field of veterinary biological products. A novel goose parvovirus strain is obtained through separation, and poultry can have neurological symptoms after being infected with the goose parvovirus strain for 2-3 days. The novel goose parvovirus is inactivated, the prepared novel goose parvovirus inactivated vaccine is good in safety, effective immune protection can be provided for ducks, and the novel goose parvovirus inactivated vaccine has good commercialization development prospects. When the novel goose parvovirus inactivated vaccine is prepared, firstly, the novel goose parvovirus is inoculated into DEF cells in the S phase to obtain virus liquid, on the basis, an oil phase and a water phase are prepared according to a proper formula proportion, and full emulsification is performed, so that the preparation efficiency of the inactivated vaccine is improved, and meanwhile, the novel goose parvovirus inactivated vaccine is prepared. The safety and the stability of the prepared vaccine are improved.

The invention discloses a goose parvovirus strain adapted to various waterfowl embryo fibroblasts as well as infectious clone and application of the goose parvovirus strain. The genome sequence of the goose parvovirus strain provided by the invention only generates three base mutations in the VP1 protein, resulting in the generation of amino acid mutations at three sites. The JNC23 shows a typical cytopathic effect in the growth of embryo fibroblasts of cherry valley ducks, Jinding ducks, Shaoxing ducks, geese and muscovy ducks. The half tissue cell infection amount of the JNC23 on the poultry embryo fibroblasts reaches 105.6 / ml to 107.6 / ml. The invention further provides a construction and rescue method of the infectious plasmid pJNC23 of the JNC23 strain, and the construction and rescue method can be used for further deeply exploring the mechanism that amino acid point mutation of the encoding protein affects NGPV replication and pathogenicity.

The application relates to a primer and a locked nucleic acid probe group for real-time fluorescent quantitative PCR detection of two strong and weak strains of goose parvovirus, wherein the strong and weak strains of goose parvovirus include MDGPV strong and weak viruses and SBDSV strong and weak viruses, and the sequences of the primer and the locked nucleic acid probe group are shown in SEQ ID NO. 1-8. The LNA-TaqMan real-time fluorescent quantitative PCR detection method established by using the primer and the LNA-TaqMan probe group can simultaneously detect, differentially diagnose and accurately quantify the strong and weak strains of goose parvovirus (duck small goose plague virus and / or duck short-beak dwarf syndrome virus strong and weak viruses), and is high in sensitivity, strong in specificity and good in repeatability.