Preparation method of identifiable inactivated vaccine for animals

A technology for inactivated vaccines and animals, applied to medical preparations containing active ingredients, pharmaceutical formulas, antibody medical components, etc., can solve problems such as re-epidemic, mutation, and residual vaccine strains

Inactive Publication Date: 2014-10-01
SHANGHAI CHUANG HONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, the safety problem of using bacteria and viruses for the production of attenuated live vaccines is that animals may have residues of vaccine strains, gene recombination, gene variation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of Porcine Pseudorabies Virus Differentiable Inactivated Vaccine:

[0023] Isolation of new epidemic strains of porcine pseudorabies: Aseptically collect brain, lung, kidney and other tissues from the dead piglets in the pig farm, mix them and cut them into pieces in a sterilized mortar. According to this method, 3 piglets were processed The diseased material was diluted 1:5 with DMEM to form a suspension, and was repeatedly frozen and thawed at -70°C for 3 times. After centrifugation at 3000r / min for 30 minutes, take the supernatant and filter it through a 0.22 μm microporous membrane, add appropriate amount of penicillin and streptomycin, inoculate it into the culture flask of BHK-21 cells that have grown into a single layer, and place it at 37 ℃ incubator to absorb for 1 hour, and then add DMEM maintenance solution to incubate for 72 hours to collect the poison. Purified and identified as pseudorabies virus.

[0024] Knockout of gE: Apply x-gal (40 μg / m...

Embodiment 2

[0027] Preparation of identifiable inactivated vaccine for swine fever:

[0028] The current vaccine for preventing swine fever is the C-strain live vaccine, which has a good immune effect, but it cannot be distinguished from the wild strain. On the basis of the C strain, the E2 gene was artificially knocked out, resulting in a deletion of 54 amino acids, that is, 693-746 of the B / C region. The culture, immunogenicity and effectiveness of the strain will not be affected after the knockout, and it can resist the attack of the virulence of classical swine fever.

[0029] The identifiable swine fever vaccine strain is inoculated into ST cells, calf testicular cells, etc. for cultivation, and the virus culture medium is harvested in 36 to 48 hours, and then 0.1% to 0.5% inactivator is added to the culture medium, shaken well, and placed 2~8℃ for 48 hours. After testing the completely inactivated virus liquid, mix the inactivated virus liquid and the freeze-dried protective agent...

Embodiment 3

[0031] Streptococcal identifiable inactivated vaccine preparation:

[0032] The upper and lower homology arms P1 and P2 of the srtA gene and their full-length sequences were amplified, the recombinant plasmid pSET4s-P1-P2 was constructed using the temperature-sensitive "suicide" plasmid pSET4s, and the plasmid was electrotransformed into wild strains In SS2 (SC21), the srtA gene deletion strain was obtained through double screening with antibiotics and temperature. The deletion mutant strain can be inherited stably without affecting the culture characteristics, and still maintains good immunogenicity.

[0033] The cultivation, inactivation and compounding of vaccines are the same as those of conventional vaccines.

[0034] The vaccine of the present invention can be used after passing the following tests: sterility test, exogenous virus test, safety test and efficacy test.

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Abstract

The invention discloses a preparation method of identifiable inactivated vaccine for animals, and relates to the technical field of animal vaccines, in a preparation process, a gene engineering method, a natural subculture method or a chemical synthesis method is adopted to reduce one or multiple segments of genes from antigenic substance to generate new properties, and the antigenic substance with new properties is prepared into the identifiable inactivated vaccine. The preparation method disclosed by the invention has the beneficial effects that after the vaccine is inoculated to an animal, an antibody can be verified by detecting the properties, by verifying whether the antibody is generated by the non-treated antigen (namely, wild virus) or by treated antigen (namely, vaccine virus), the antibodies generated by the vaccine and the wild virus can be distinguished to diagnose whether the animal is infected with wild virus strain, so as to eliminate and remove animals with virus to eliminate diseases, thereby being beneficial to eliminate diseases in farms.

Description

technical field [0001] The invention relates to the technical field of animal vaccines, in particular to a method for preparing an identifiable inactivated vaccine for animals. Background technique [0002] Now that there are more and more animal diseases, vaccines have become the main tool for disease prevention and control. Live attenuated vaccines play a very important role in preventing the occurrence of infectious diseases and controlling the outbreak of infectious diseases. However, the safety problem of using bacteria and viruses for the production of attenuated live vaccines is that animals may have residues of vaccine strains, gene recombination, gene variation, and strong virulence after long-term use of attenuated live vaccines, causing the disease to become popular again. It is not conducive to the purification and removal of infectious diseases. [0003] Animal vaccines play a vital role in the prevention of animal diseases. Many infectious diseases at home and...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/39
Inventor 赖志高俊锋马晶晶吴碧清
Owner SHANGHAI CHUANG HONG BIOTECH
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