Method for preparing heat-resisting attenuated virus live vaccine for goatpox by using BHK21-C13 passage cell

Abstract

The invention discloses a method for preparing a heat-resisting attenuated virus live vaccine for goatpox by using a BHK21-C13 passage cell. Based on the existing attenuated virus AV41 for goatpox, which has excellent immunogenicity, the method comprises the following steps: culturing 25-28th generations of virus solution with a heterologous BHK21 cell, and then adding an appropriate heat-resisting freeze-drying protective additive to obtain the heat-resisting attenuated virus live vaccine for goatpox. The passage cell vaccine used by the method is superior to the primary cell vaccine in the aspects of virus yield, homogeneity, purity and the like, not only guarantees the effective level of the vaccine and has a better heat-resisting protection effect, but also can ensure the safety of a purebred pregnant goat. As the existing BHK21 cell adopts a mature suspension culture process, the exploration of various parameters of adherent culture lays a solid foundation for the next promotion of suspension process.

Description

technical field [0001] The invention belongs to the preparation method of vaccines in the field of biomedicine, and in particular relates to a method for preparing goat pox serum-free attenuated live vaccine virus liquid with BHK21-C13 subcultured cells, and adding a heat-resistant protective agent to the product to obtain a heat-resistant type for veterinary use Goat pox attenuated live vaccine method. Background technique [0002] Goat pox is a highly contagious disease of goats caused by goat pox virus (GPV). Sick sheep manifested as elevated body temperature, papules or nodules, blisters, visceral pox lesions on the skin and mucous membranes of hairless or less hairy parts, abortion in pregnant ewes, and high mortality. Goat pox is a legally notifiable infectious disease stipulated by the World Organization for Animal Health (OIE), and it is listed as a first-class animal disease in my country. [0003] The existing vaccines for preventing goat pox are mainly attenuate...

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Problems solved by technology

The existing heat-resistant protective agent formula one (patent document CN104800842A) is: trehalose 50g / L, skimmed milk powder 20g / L, polyvinylpyrrolidone 20g / L, gelatin 10g / L, arginine 1.5g / L, vitamin C 2.5g / L; Formula 1 was used for goat pox sheep testicular primary cell live vaccine, but the cost was high, and its heat-resistant effect was not verified; Heat-resistant protective agent formula 2 (porcine pseudorabies live vaccine heat-resistant protective agent and frozen Drying Technology Research—Wang Ying: Gansu Agricultural University 2014 Master's Degree Thesis, Chapter 3 Screening of Porcine Pseudorabies Virus Heat-resistant Protective Agents, page 38) (W / V): sucrose 15.2%, enzymatic casein 7.6% , 4.0% gelatin, 1.0% trehalose; 2.0% bovine serum albumin, 2.0% arginine hydrochloride, 2% sodium glutamate, 2.0% sorbitol, and the rest are water for injection; formula two can be used for other viruses ( For example, attenuated live vaccines such as pseudorabies virus, etc., have not been used for goat pox attenuated live vaccines, and it is not yet known whether they are applicable

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