11 results about "Tembusu virus" patented technology

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 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 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 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 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 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.