68 results about "Totivirus" patented technology

The invention discloses a large-scale preparation method for foot-and-mouth disease totivirus marked vaccine with high yield, high purity and high safety and a product thereof. The method comprises the following steps: a)collecting a virus solution; b)performing deep filtration on a membrane, performing ultrafiltration and performing enzymolysis on nuclease; c)purifying through a strong anion exchange adsorption bed or an adsorption film; d)depositing by PEG, extracting by chloroform-isoamyl aleohl; e)inactivating; F)performing density gradient centrifugation on an inactivation liquid through cane sugar and purifying; g)performing ultrafiltration dialysis and aseptic filtration; and h)reserving a stock solution or emulsifying. The provided foot-and-mouth disease totivirus marked vaccine antigen is uniform and complete foot-and-mouth virus particle, The vaccine is injected into body, so animal infection and immunization can be completely distinguished, does not contain foot-and-mouth disease virus non-structural protein and other virus particle, and does not contain animal-based foreign protein, polypeptide and oligopeptides, animal latent anaphylactic reaction, carcinogenesis and latent risk such as mad cow disease for causing animal infectious diseases due to vaccine injection can be effectively reduced, and the vaccine has no influence on animal food safety and trade.

The invention discloses a large-scale full-suspension production method of a porcine circovirus type 2. The inventors of the invention domesticate a porcine kidney cell adaptable to large-scale serum-free full-suspension culture, which is named as sPK15-YP, is collected in the China General Microbiological Culture Collection Center and has the culture collection number of CGMCC NO.13846. The sPK15-YP cell adaptable to full-suspension culture is used for achieving serum-free large-scale culture of the porcine circovirus type 2 (PCV2), so that the conventional spinner bottle culture technology is replaced, the human resource is reduced, the product quality is improved, and the bottleneck that the virus content is low during PCV2 full-virus culture is solved; by a full-suspension sPK15-YP cell technology, a high-potency PCV2 semifinished product is prepared; by a hollow fiber method, a PCV2 virus culture solution is concentrated and purified to obtain a more pure PCV2 virus concentrated antigen. By the large-scale full-suspension production method, a solid foundation is laid for studying a PCV2 multivalent vaccine, the dosage of the vaccine is reduced, the stress of a swine herd is reduced and the immune level of the swine herd is enhanced.

The invention provides a lyophilized inactivated Japanese encephalitis vaccine, comprising: (a) inactivated Japanese encephalitis totivirus; (b) stabilizer, which comprises maltose, lactose, and optionally mycose, mannite, sorbitol and / or amino acid; and optionally (c) buffer, surfactant, isotonic regulator and / or chelating agent.

An external culture system of hepatitis C totivirus for culturing complete multiplicative HCV virus by the molecular biology and gene recombination technique is disclosed. Its culture steps include amplifying the full-length genom containing 98 nucleotides after 3' PolyA in the serum of hepatitis C patient; artificial site-directed mutation to NS5A and NS5B in HCV genon; inserting the expression box of marker gene IRES-GFP at NS5B3' terminal in the mutated HCV genom; transfecting sensitive cell and culturing to obtain progeny HCV virus with infection activity.

The invention provides a vaccine composition, comprising porcine pseudorabies virus gE gene-deleted strain of immunizing dose, or an attenuated live totivirus antigen or inactivated totivirus antigen of the culture thereof. The vaccine composition is effective in inducing generation of antibodies to provide effective protection for swine and can be used as a marker vaccine to effectively distinguish wild strains and vaccine strains.

The invention relates to a rabies virus monoclonal antibody with neutralization activity and capability of specifically recognizing rabies virus glycoprotein G. The monoclonal antibody is authenticated to be IgG1-subtype. According to the rabies virus monoclonal antibody disclosed by the invention, rabies virus inactivation totivirus is taken as an immunogen for immunizing a mouse, an antibody with neutralization activity is screened, the neutralization titer is high, the secretion of hybridoma for the antibody is stable, and the antibody is easy to store; and an in-vitro preparation link for glycoprotein is avoided, and innovativeness is achieved. A colloidal gold test paper card or test paper strip prepared from the antibody is high in test sensitivity and capable of achieving 0.03 IU / mL, capable of being used for test on human rabies virus or quality control in a vaccine production process, and high in application value.

The invention discloses a kind of bluetongue virus (BTV) NS2 protein monoclonal antibody BTV-4D4, a B cell epitope recognized thereby and applications. BALB / c mice are subjected to totivirus immunization by utilization of bluetongue virus serotype 8 type (BTV8), splenic lymphocytes of the mice after immunization and SP2 / 0 cells are fused. A hybridoma cell strain BTV-4D4 secreting anti-BTV-NS2 protein monoclonal antibodies steadily is obtained after screening with BTV 8 as a coating antigen through an indirect ELISA method. The monoclonal antibodies secreted from the hybridoma cell strain and the BTV-NS2 protein are subjected to a specific reaction. The B cell epitope recognized by BTV-4D4 is 149RSNYDV154 after identification by utilization of a phage display technology. The monoclonal antibody and the BTV-NS2 protein B cell epitope recognized by the monoclonal antibody can be used for diagnosis or prevention of bluetongue virus infection.

The invention discloses an application of a soluble I-type duck hepatitis virus 3D protein to preparation of an ELISA reagent and an ELISA kit. The ELISA kit comprises a soluble I-type duck hepatitis virus 3D protein coated reaction plate, an enzyme-labeled antibody, a developing solution and a stop solution. When being used for detecting I-type duck viral hepatitis antibodies, the ELISA kit is strong in specificity, good in reproducibility and high in sensitivity, and has a higher agreement rate than the ELISA kits taking totivirus as coating antigen in the aspect of detection, so that the ELISA kit can be used for the detection of clinical samples.

The invention discloses a preparation method and application of an anti-PCV2 monoclonal antibody. The anti-PCV2 monoclonal antibody, and DNA sequences and amino acid sequences of the heavy chain variable region and the light chain variable region of the antibody are disclosed. The preparation method of the antibody is designed. The antibody is applied in an antigen / antibody detection kit, antigen / antibody immunochromatography test paper, IFA, IPMA and a PCV2 or PCV2 Cap immunoaffinity column. The antibody can simultaneously recognize two major epidemic subtypes which are PCV2a and PCV2b, and has good reactogenicity with PCV2 totivirus and PCV2 Cap protein, but is free of cross reaction with PCV1 and PCV3 subtypes and porcine viruses, and lays a foundation for the study of etiology and pathogenesis of PCV2 and clinical detection of PCV2 pathogens.

The invention provides a cell line highly sensitive to a porcine circovirus type 2 (PCV2) and discloses a preparation method of the cell line. After the PCV2 is adopted to infect PK-15 cells, expression of 3-hydroxyl-3-HMG-CoAreductase (HMGCR) is obviously changed; and after HMGCR genes are subjected to silence, TCID50 (tissue culture infectious dose 50) of the PCV2 reaches 108.5 / ml. An HMGCR gene silence cell line can be obtained by RNA (ribonucleic acid) interfering technology, and the cell line highly sensitive to the PCV2 can be finally obtained through screening. By utilizing HMGCR gene silence cells prepared by the method to culture the PCV2, high-titer viruses can be obtained, and studying on pathogenesis of the PCV2, virus culturing and development and industrial production of totivirus inactivated vaccines are facilitated.