22 results about "Tembusu virus" patented technology

The invention discloses application of GSK2606414 in preparation of a medicine for inhibiting duck tembusu virus infection, and belongs to the technical field of veterinary biological products. The research finds that the GSK2606414 can effectively inhibit the replication of the duck tembusu virus for the first time, provides a new therapeutic drug for the prevention and treatment of the duck tembusu virus, and is of great significance to the actual production of the poultry breeding industry.

The invention belongs to the field of biology, and relates to a broad-spectrum monoclonal antibody aiming at avian influenza virus M1 protein and application of the broad-spectrum monoclonal antibody, the monoclonal antibody has specific reaction with A549 cells infected by avian influenza, has no specific reaction with Newcastle disease virus, duck tembusu virus, goose astrovirus and infectious laryngitis virus, has good specificity, and can be used for preparing the broad-spectrum monoclonal antibody. The 5G9 monoclonal antibody has good reactivity with A549 cells infected by H1-H11 subtype AIV, has good broad spectrum, can be used for detecting M1 protein sub-localization after the AIV infected cells are detected by using the 5G9 monoclonal antibody, and can be used for indicating the infection process of the AIV.

The invention discloses a DNA vaccine plasmid of a tembusu virus as well as a construction method and application of the DNA vaccine plasmid, and relates to the technical field of molecular biology. The DNA vaccine plasmid of the tembusu virus comprises an expression vector pcDNA3.1-prME containing a 5AA-STING gene segment, and a DNA vaccine plasmid of the tembusu virus, the 5AA-STING gene segment is a gene segment formed by introducing a coding sequence of five amino acids at the N end of the tembusu virus NS1 on an STING gene segment. The invention further provides a construction method and application of the DNA vaccine plasmid of the tembusu virus, the DNA vaccine plasmid of the tembusu virus is favorable for efficiently assembling into a subvirion with a stronger immune effect, and the DNA vaccine plasmid of the tembusu virus can co-express an immune stimulant STING molecule while expressing the structural protein of the tembusu virus to be assembled into the subvirion, so that the DNA vaccine plasmid of the tembusu virus has a good application prospect. The polypeptide is a key molecule for recognizing a double-stranded DNA signal channel in cells, and the immune protection effect is further enhanced.

This invention discloses a primer set for multiplex PCR detection of duck plague virus (DTMUV), duck Tembusu virus (DPV), and Riemerella anatipestifer, a kit containing this primer set, and their applications. This invention optimizes and establishes a multiplex PCR detection method for DTMUV, DPV, and Riemerella anatipestifer. Specificity results show that the multiplex PCR detection method established in this invention can only amplify specific bands for DTMUV, DPV, and RA, while no amplification bands are found in PCR amplification products using other pathogen DNA or cDNA as templates, resulting in negative results. Specificity test results demonstrate that the multiplex PCR detection method established in this invention has good specificity. Sensitivity test results show that the minimum detection limit for DTMUV cDNA is 9.855 pg, for RA DNA is 0.9 pg, and for DPV DNA is 0.66 pg. This invention provides an effective technical means for the diagnosis of duck Tembusu virus disease, duck plague, and Riemerella anatipestifer disease.

The application discloses a highly immature duck Tembusu virus, a culture method and application thereof, and belongs to the technical field of biotechnology.According to the method for culturing the highly immature duck Tembusu virus based on the LOVO cell line provided in the application, the duck Tembusu virus obtained by culture loses pathogenicity, the application provides a highly potential candidate strain for developing safe and effective duck Tembusu virus vaccines, opens up a new way for safely producing high-quality diagnostic antigens and vaccine antigens, and significantly reduces the biosafety risk of related research.

The present invention discloses a duck Tembusu virus CHN-YC and a dual inactivated vaccine thereof and a preparation method. The deposit number of CHN-YC is CCTCC NO: V202371. The vaccine comprises an inactivated duck Tembusu virus CHN-YC and an inactivated novel duck reovirus QR-China / 2020. The present invention obtains a highly pathogenic duck Tembusu virus strain, and the inoculation dose of the virus to 5-day-old SPF ducks is 10 7 TCID 50 The lethality rate reaches 55%. Furthermore, the strain has strong immunogenicity and is suitable for use as a vaccine strain. The dual inactivated vaccine of the present invention is relatively stable, enabling ducks injected with the vaccine to produce high levels of serum neutralizing antibodies, thereby achieving better immunization against the novel duck reovirus and duck Tembusu virus. Consequently, ducks injected with the dual inactivated vaccine are less susceptible to diseases caused by the two viruses, allowing for simultaneous immunization against two diseases.

The present invention discloses an HSPA8 protein that is resistant to duck Tembusu virus and its application. The present invention constructs a prokaryotic expression vector for expressing HSPA8, optimizes protein expression conditions, and expresses and purifies soluble, endotoxin-free HSPA8 proteins, including HSPA8-His protein and GST-HSPA8 protein. In vitro antiviral experiments demonstrate that the HSPA8 protein of the present invention has a good antiviral effect; in vivo antiviral experiments demonstrate that the HSPA8 protein can significantly reduce the pathogenicity of DTMUV infection, prevent viral proliferation in vivo, reduce weight loss in mice caused by viral infection, and reduce mortality in mice caused by viral infection. Therefore, the HSPA8 protein of the present invention has significant resistance to DTMUV infection and can be used to prepare drugs against duck Tembusu virus, providing a new strategy for the prevention and control of duck Tembusu virus.

The invention discloses a highly immature duck tembusu virus as well as a culture method and application thereof, and belongs to the technical field of biology, and the duck tembusu virus cultured according to a method for proliferation culture of the highly immature duck tembusu virus based on an LOVO cell line loses pathogenicity. The invention provides a potential candidate strain for developing a safe and effective duck tembusu virus vaccine, opens up a new way for safely producing high-quality diagnostic antigens and vaccine antigens, and remarkably reduces the biological safety risk of related researches.

The invention belongs to the field of cell biology, and relates to a shelduck fibroblast cell line which is finally obtained by performing primary culture, screening and optimized subculture on shelduck embryos serving as materials. The fibroblasts cultured by the method are slender, fusiform, triangular and irregular, and can be continuously subcultured for at least 82 generations; the shelduck fibroblast cell line is used for culturing waterfowl viruses such as duck plague virus, novel duck reovirus, duck tembusu virus, duck parvovirus and the like for the first time; compared with the existing duck-derived fibroblast culture, the duck-derived fibroblast culture medium has the advantages of high virus content and wide application range. The shelduck fibroblast cell line provided by the invention can provide a large amount of high-quality materials for scientific researches of cell engineering, molecular biology, immunology and the like; the method is used for culturing, separating and identifying poultry viruses, provides raw materials for development and production of poultry attenuated vaccines and inactivated vaccines, can shorten the production period and reduce the production cost, and has certain practical significance for large-scale production of vaccines.

The application discloses a duck Tembusu virus attenuated strain, and a nucleotide sequence of the duck Tembusu virus attenuated strain is shown as SEQ ID NO:1.The duck Tembusu virus attenuated strain can immunize ducklings and make the ducklings produce effective duck Tembusu virus antibodies, the protection rate of the ducklings reaches 100%, meanwhile, the weight growth of the ducklings is not affected, and there is no virus discharge, death and viremia, and the duck Tembusu virus is not detected in main organs of the ducklings.The duck Tembusu virus attenuated strain can also be used as a live vector to embed H5N6 subtype avian influenza virus antigen epitopes, and further construct a live vector vaccine strain, and the live vector vaccine strain can immunize ducklings and effectively reduce the damage of H5N6 subtype avian influenza virus to the ducklings.

The invention discloses a triple fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection kit and a detection method for simultaneously detecting an avian influenza virus, a duck tembusu virus and a novel duck reovirus, specific primers and TaqMan probes are respectively designed aiming at gene conserved regions of the three viruses, reaction conditions are optimized, and the specific primers and TaqMan probes can be used for simultaneously detecting the avian influenza virus, the duck tembusu virus and the novel duck reovirus. A TaqMan triple real-time fluorescent quantitative RT-PCR diagnosis method capable of simultaneously detecting three viruses is established and is applied to clinical sample detection. Results show that the established triple fluorescence RT-PCR method has high sensitivity and strong specificity, can meet the requirements of high throughput and rapid detection, and has high application value.

The invention discloses an epitope tandem recombinant protein X and application thereof in duck tembusu virus blocking ELISA (enzyme-linked immuno sorbent assay) detection, and belongs to the technical field of molecules. The antigen epitopes A, B, C and D obtained through screening are connected in series, and the epitope series recombinant protein X is obtained. A duck tembusu virus blocking ELISA detection method is established by using the epitope tandem recombinant protein X as a coating antigen protein, the epitope tandem recombinant protein X as a coating antigen is compared with ELISA using A protein, A and B tandem protein and A, B and C tandem protein as coating antigens, and the result shows that the epitope tandem recombinant protein X as the coating antigen can be compared with the ELISA using A protein, A and B tandem protein and A, B and C tandem protein as the coating antigen. The epitope tandem recombinant protein X is taken as a coating antigen, so that the specificity is relatively high.

The invention discloses an epidemic disease prevention, detection and purification method for Sansui duck breeding, which is used for daily epidemic disease prevention and control of commercial duck farms and breeding duck farms of Sansui ducks, and comprises the following steps: S1, epidemic disease detection and screening aiming at duck tembusu virus disease, riemerella anatipestifer disease and duck poultry pathogenic colibacillosis; s2, obtaining a detection result in the step S1 for isolation; s3, breeding monitoring and activity track analysis are checked for infection prevention; s4, disinfecting the functional area during a non-epidemic period, collecting an environmental swab, and laying an infection prevention area; s5, after the S3 and the S4 are executed, the S1 is executed regularly, and continuous monitoring implementation is carried out for more than six months; and S6, when the positive individuals appear in the S5, repeating the steps S3-S5 until no positive individuals exist in the continuous monitoring period. Compared with the prior art, the epidemic disease prevention, detection and purification method for Sansui duck breeding has the advantages of being good in purification, prevention and control effect and capable of synchronously aiming at DTMUV, RA and APEC epidemic diseases.

The application discloses a colloidal gold immunochromatography test strip for rapidly detecting avian Tembusu virus, a preparation method and application thereof, and relates to the field of virus detection. The mononuclear hybridoma cell of the anti-DTMUV E protein is obtained by fusing myeloma SP2 / 0 cells with immune Balb / c mouse spleen cells. The specific antibody for the DTMUV E protein is obtained by hybridizing the mononuclear antibody (mAb) with the previously prepared mouse polyclonal antibody. The immunochromatography test strip is prepared by taking the colloidal gold-mAb combination and the capture antibody as targets. The test strip is high in specificity, can accurately detect the DTMUV, has a detection lower limit of 26.1718 copies, is good in repeatability, can detect various complex samples, and is intuitive in results. The GICA test strip established in the application is a rapid, specific and stable on-site preliminary diagnosis tool for the DTMUV related symptoms, and is favorable for the monitoring and prevention and control of the DTMUV in resource-poor areas.

The invention belongs to the technical field of virus detection, and particularly relates to a PCR (Polymerase Chain Reaction) detection kit for four vertical transmission viruses of waterfowl and application of the PCR detection kit. The kit can rapidly, specifically and simultaneously amplify and diagnose duck hepatitis virus type I (DHV-1), duck Tembusu virus DTMUV, Muscovy duck reovirus MDRV and duck astrovirus DAstV, has high specificity and sensitivity, can accurately and rapidly identify DHV-1, DTMUV, MDRV and DAstV, can complete fluorescent quantitative PCR detection in about 50 minutes, and is simple to operate and practical. The detection period is shortened to 1.5 hours from 6-8 hours of a traditional method, the sensitivity is improved by 10-1000 times, the early diagnosis capability of epidemic diseases is remarkably improved, and the epidemic disease misjudgment rate of a farm is obviously reduced. Abuse of vaccines and drugs is reduced, and sustainable development of the poultry breeding industry is promoted.

This invention discloses a linearized mutant plasmid of Tembusu virus SLII, a recombinant virus, and its applications, belonging to the field of molecular biology. Using the pACNR CQW1-Intron infectious clone as a backbone, the SLII stem-loop structure within the 3′UTR Domain-I region was linearized to construct the SLII linearized mutant infectious clone pACNR-CQW1-SLIILiner, which was then rescued to obtain the recombinant virus CQW1-SLIILiner. This recombinant virus exhibits limited replication capacity at the in vitro cellular level and displays a significantly attenuated phenotype in animal models. It can also induce specific neutralizing antibodies and cellular immune responses, providing significant protection against virulent strains. Therefore, the recombinant virus of this invention possesses advantages such as clear molecular genetic markers, high safety, good immunogenicity, and strong genetic stability, and can be applied to the preparation of live attenuated vaccines or related biological products for the prevention of TMUV infection.

The invention discloses a polypeptide for inhibiting duck tembusu virus replication and application of the polypeptide as a medicine for inhibiting duck tembusu virus replication, and the polypeptide is scorpion active polypeptide Eval418. When the scorpion active polypeptide Eval418 provided by the invention is used as a drug for inhibiting duck tembusu virus replication, the scorpion active polypeptide Eval418 can efficiently inhibit DTMUV replication at a cellular level with low toxicity, and the mechanism of the scorpion active polypeptide Eval418 relates to a plurality of early links such as direct destruction of virus particles, blocking of virus attachment and invasion and the like. According to the technical scheme, a new candidate is provided for prevention and treatment of the DTMUV.

The invention belongs to the technical field of biology, and discloses an application of emodin-8-glucoside in resisting tembusu virus. An in-vitro experiment is carried out by using a recombinant duck tembusu virus stably expressing EGFP (Enhanced Green Fluorescent Protein), the emodin-8-glucoside is proved to be capable of efficiently inhibiting virus replication, the median effective concentration (EC50) of the emodin-8-glucoside is as low as 1.935 mu M, the median cytotoxic concentration (CC50) of the emodin-8-glucoside is as high as 511.3 mu M, and the calculated therapeutic index (SI) is 264.24, so that the compound has the outstanding characteristics of high efficiency and low toxicity, and can be used for preparing a medicine for preventing and treating the virus replication. The clinical application safety is high; duck infection model experiments prove that the emodin-8-glucoside can effectively inhibit the duplication of DTMUV in duck bodies, and has a definite treatment effect; meanwhile, the invention also provides a medicine, a feed and a feed additive containing the emodin-8-glucoside.

The invention is applicable to the field of immunodetection, and provides a double-antibody sandwich detection kit for duck tembusu virus as well as a detection method and application thereof, and the double-antibody sandwich detection kit comprises the steps of preparation of a mouse monoclonal antibody, preparation of a duck polyclonal antibody, enzyme-linked immunosorbent assay (ELISA) detection procedure and the like. Wherein the capture antibody is a mouse monoclonal antibody of an anti-TMUV-prM protein, and is derived from a hybridoma cell strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.45754; the detection antibody is an anti-TMUV duck polyclonal antibody. The invention provides a kit for detecting TMUV (TetramethylUV) prM protein and / or TMUV and a detection method. The kit and the method have excellent specificity, sensitivity and repeatability, can be used for rapidly and accurately detecting TMUV, and are suitable for being popularized in clinical detection.

The invention provides a chicken tembusu virus antigen protein, a subunit vaccine and application thereof, belongs to the technical field of vaccine preparation, and solves the technical problem that a chicken tembusu virus subunit vaccine special for chicken does not exist in the market at present. According to the chicken tembusu virus antigen protein provided by the invention, the amino acid sequence of the antigen protein is selected from E-GL protein as shown in SEQ ID NO: 3 or E-HA protein as shown in SEQ ID NO: 6. The method can be applied to challenge protection of chicken tembusu virus strains, and has the advantages of high safety, strong effectiveness, high expression quantity and great reduction of production cost.

The invention discloses an RT-ERA-CRISPR / Cas12a composition for detecting a duck tembusu virus. The RT-ERA-CRISPR / Cas12a composition comprises an RT-ERA primer group and crRNA (Complementary Ribonucleic Acid), the RT-ERA primer group comprises an upstream primer with a nucleotide sequence as shown in SEQ ID NO: 1 and a downstream primer with a nucleotide sequence as shown in SEQ ID NO: 2, or an upstream primer 1 with a nucleotide sequence as shown in SEQ ID NO: 11 and a downstream primer 1 with a nucleotide sequence as shown in SEQ ID NO: 12; the nucleotide sequence of the crRNA is as shown in SEQ ID NO: 13 or SEQ ID NO: 14. The RT-ERA-CRISPR / Cas12a composition can be used for detecting 1 copy / mu L of duck tembusu virus RNA at 42 DEG C within 15 minutes, and the method is simple, short in time consumption and high in sensitivity.

The application relates to the technical field of molecular biology, and particularly discloses a mutant duck Tembusu virus strain, a construction method and application thereof. A mutant plasmid EDII+III E89G V312A of site-directed mutation E gene is constructed through specific primer combination, and then a mutant duck Tembusu virus strain rDTMUV EDII+III E89G V312A is constructed. The strain can provide a good platform for preparation of subsequent anti-duck Tembusu virus drugs and vaccines.