40results about How to "High poison price" patented technology

The invention discloses mass production of inactivated vaccine of porcine circovirus II (PCV2) and a preparation method thereof. The preparation method comprises the following technical steps: (1) high-density culture of cells for vaccine preparation; (2) reproduction of venom for vaccine preparation; and (3) addition of adjuvant to prepare the inactivated vaccine. Compared with the prior art, the invention has the advantages that the virus yield is high, the virus titer is high, the production scale is large, the yield of a single batch is high, the production cost is relatively low, the product quality is high and stable, the operation is convenient, the operation space is small, the technological parameters are controlled accurately and the like. The inactivated vaccine has high safety, can induce pig bodies to generate immune protection and fully satisfies the national biological product standards.

The invention discloses a culture method of porcine pseudorabies virus; hamster kidney cells (BHK-21) are used as a cell source for culturing the porcine pseudorabies virus, and while cell passage, the virus is inoculated, so that multiplication culture of the virus is realized. The method has the characteristics of simple technology, high increment, high yield and low cost, and a vaccine prepared by use of the porcine pseudorabies virus cultured by the method has a complete preventive effect on the porcine pseudorabies virus, and has good social benefits and application prospects.

The invention belongs to the technical field of veterinary biological products, and particularly relates to a BHK-21 cell line BHK-21-BP-2 clone and application thereof in full suspension cell culture. The provided BHK-21-BP-2 clone is preserved in the China general microbiological culture collection center, and the preservation number is CGMCC No. 17588. After the clone is inoculated with pseudorabies viruses during full suspension cell culture, the clone has rapid cytopathy, the virulence values of generated viruses are high, and the viruses can be collected after 24-48 hours; at the time ofvirus collection, only supernatant needs to be collected by centrifugation without repeated freezing and thawing, and the production process is simple. The total protein content of a produced virus antigen solution is low, the integrality and quality of obtained effective viral antigens are high, the viruses are excellent in immunogenicity and safety, and a prepared pseudorabies virus vaccine hasthe advantages of high stability, small difference between batches, a good immune effect and high safety.

The invention provides a mutated PCV2 (Porcine circovirus Type 2) virus free from being degraded by ubiquitination proteasome, and a preparation method and application thereof. Three loci (164K, 179Kand 191K) of porcine circovirus 2 type (PCV2, GenBank and No. EU366323) strains Cap are mutated, the mutated PCV2 can not be subjected to MKRN1-mediated ubiquitination degrading, in addition, high speed copying can be carried out through infectious clone, and virus price is high than virus price of wild-type PCV2. Through mutated viruses obtained by the infectious clone, the pathogenic mechanism of the PCV2 can be more favorably researched, and a potential control measure is provided for PCV2 outbreak.

The invention provides a chicken newcastle disease virus inactivated vaccine preparation method. According to the present invention, inoculation with chick embryo fibroblasts is used to replace inoculation with chick embryo allantocherion to breed chicken newcastle disease viruses, such that the vaccine preparation cost is saved, the quality difference between the batches is reduced, the yield is high, and the virus titer and the vaccine quality are improved; the vaccine is emulsified with the homogenizer, such that the prepared vaccine has advantages of low viscosity, small particle size and easy absorption compared to the traditional water-in-oil dosage form; and any surfactant components are not required to be added during the vaccine preparation process through the selected adjuvant, the prepared vaccine has good safety, and the side effect due to the difficult absorption of the traditional oil adjuvant is avoided.

The invention provides a novel method for preparing porcine circus-virus 2 type, wherein inoculated PK-15 cells are treated by D-glucosamine with different concentration gradients, and is screened by IFA; the method not only makes the OK-15 cells normal grow, but also substantially raises PVC2 virus titer; the PVC2 virus titer acquired by the method is not less than 105TCID50, and a method for propagating and operating PVC 2 by screened proper D-glucosamine is used for researching a PVC2 vaccine and a relative diagnostic reagent, and preparing the products.

The invention relates to a method for producing swine fever vaccines, which comprises the following technical steps: (1) cloning and identifying hog cholera lapinized virus strain; (2) propagating seed viruses of the cloned strain; (3) propagating productive cells; (4) propagating productive virus solution; and (5) distributing vaccines, filling and freeze-drying. The method of the invention has the characteristics of simple and stable production process, high virus immunogenicity and genetic stability, small exogenous virus pollution risks, convenient examination, low production cost and the like. The swine fever vaccines produced by the method have high genetic stability, purity and immunogenicity.

The invention discloses two siRNA sequences and target gene sequences thereof. Two target spots of si-LNC_002015 gene in Vero-E6 culture cells are interfered through siRNA, so that the PEDV RNA copy number and the protein expression quantity in the Vero-E6 culture cells are obviously increased compared with those of a control group, and the virus titer is increased.

The invention discloses a method for constructing a muscovy duck reovirus (MDRV) reverse genetic system. Viral genome RNA is extracted from cell culture poison containing the muscovy duck reovirus; adesigned specific primer and the viral genome RNA serving as a template are utilized to perform RT-PCR amplification on gene segments of an MDRV complete sequence, enzyme digestion is performed on target gene segments to connect the target gene segments into 10 gene segments of the MDRV, the 10 gene segments are connected with a carrier, and positive bacterial colonies are picked to perform sequencing and verification; positive plasmids of the 10 segments of the MDRV are mixed according to the proportion to perform cotransfection on single-layer 293T cells. After passage is performed on the cells for 15th generation, cytopathy becomes stable, and MDRV recombinant virus can be obtained.

The invention relates to the field of cells and biological products, in particular to a serum-free full-suspension cell culture method for Newcastle disease VII type attenuated strain. In the method provided by the invention, the Newcastle disease virus strain is a Newcastle disease gene VII type virulent strain attenuated by F gene locus mutation. According to the method disclosed by the invention, a serum-free full-suspension cell culture is adopted, so that the method is not influenced by insufficient hatching eggs supply or uncontrollable factors; the method disclosed by the invention has the advantages of easy operation, large virus preparation amount and high virus content, and overcomes the defect that the supply of vaccines on the market is influenced by vaccine production stagnation caused by influence on virus reproduction due to insufficient supply of hatching eggs in virus chick embryo culture.

The invention relates to an animal vaccine, and particularly discloses a porcine epidemic diarrhea vaccine strain and a vaccine containing the porcine epidemic diarrhea vaccine strain. The porcine epidemic diarrhea vaccine strain adopts porcine epidemic diarrhea virus attenuated by separation and purification as virus seed, and is freeze-dried by suitable protecting agent; the virus is injected topigs by muscle injection, thus high neutralizing antibody level is generated and passive protection is provided for suckling pigs; the vaccine has the advantages of simple preparation technique, stable property, and good immunogenicity; besides, the vaccine can effectively prevent porcine epidemic diarrhea virus infection.

The invention provides a duck virus hepatitis virus, a duck virus hepatitis inactivated vaccine and a preparation method of the duck virus hepatitis inactivated vaccine. The duck virus hepatitis virus is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.7804. The duck virus hepatitis virus has strong toxicity, good immunogenicity, and high toxicity evaluation, and the vaccine prepared by the duck virus hepatitis virus has good safety and long immune protection period.

The invention relates to a duck short beak dwarf syndrome gene engineering subunit inactivated vaccine and a preparation method thereof. Aiming at a variant strain, the VP2 gene of the SBDS-GPV JS01 strain is subjected to codon optimization and is cloned to a baculovirus vector, so that a recombinant baculovirus rBac-JS01VP2 strain is successfully constructed; insect cell full suspension (or spinner bottle) culture is adopted, the technology is mature, and production transformation and step-by-step amplification culture are facilitated; compared with traditional duck embryo or duck embryo fibroblast culture, the virus titer is high, the quality is stable, the production period is short, and the cost is low; compared with an antibody product, the protection rate is high, the duration time of a maternal antibody is long, only female ducks are immunized, and labor and cost are saved; serum or egg source miscellaneous protein does not exist, and the side reaction is extremely low; the expressed VP2 protein can be assembled into virus-like particles (VLPs), and the immunogenicity is good; the expression quantity is high (when the virus titer is 8.0 Log10TCID50/ml, the VP2 protein expression quantity reaches 50-100 mg/L), the antibody level is high, and the lasting time is long; the shelf life of the product is 18 months, and the immune period is 6 months; and the ELISA antibody detection method is used for screening susceptible ducks and evaluating efficacy.