Method for preparing live vaccines of hog cholera and product thereof

A technology of swine fever live vaccine and vaccine strain, which is applied to medical preparations containing active ingredients, pharmaceutical formulas, inactivation/attenuation, etc., can solve the problem that the results of virus testing are easily affected by individual differences in rabbits and weather conditions, and cannot Accurate quantification, long test period and other issues, to achieve the effect of strong venom attack protection from swine fever, simple and stable production process, and small quality difference

Inactive Publication Date: 2010-11-10
武华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the outstanding shortcomings of this method are: the rabbit fever method is a non-specific reaction, and the poison cannot be accurately quantified, and the results of the poison test are easily affected by individual differences in rabbits and weather conditions, and the test period is long, time-consuming and laborious.

Method used

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  • Method for preparing live vaccines of hog cholera and product thereof
  • Method for preparing live vaccines of hog cholera and product thereof
  • Method for preparing live vaccines of hog cholera and product thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Screening and identification of porcine cell lines

[0028] Inoculate attenuated hog cholera virus into different pig-derived cell lines, culture at 37°C for 4 to 5 days, harvest the virus, pass the harvested virus, and use immunofluorescence method to detect the viruses inoculated with different pig-derived cell lines , The test results show that the attenuated hog cholera virus can proliferate on PT cell line, MPK cell line, SK6 cell line, PK 2a cell line, CPK cell line, and RKC cell line. The present invention uses cell lines (PT, MPK, SK6, PK 2a, CPK, RKC) to cultivate the attenuated virus of classical swine fever, harvest the cell virus liquid, and determine the virus value of the proliferating virus suspension by immunofluorescence method. The virus liquid is added with a freeze-dried protective agent and antibiotics to prepare vaccines and freeze-dried to prepare a swine fever cell vaccine.

Embodiment 2

[0029] Example 2 Method for producing swine fever live vaccine with cell line

[0030] (1) Subculture and culture of cells for seedling production: PT cell line is digested and subcultured by trypsin-EDTA cell dispersion, and is subcultured one to five. Continue culturing with cell growth solution at 37°C to form a good monolayer, which is used for continued passage or virus inoculation;

[0031] (2) Propagation of cytotoxic species: Use cell maintenance solution to prepare a 0.3% virus suspension from fresh spleen venom, inoculate a monolayer of well-growing PT cell line, and continue culturing at 37°C. The cell culture venom was harvested every 5 days as the virus seed for production; identification of cytotoxic species: the identification of cytotoxic species fully meets the standard of the attenuated strain of classical swine fever lapinization, and it has no side effects on pigs. The cytotoxic species contains more than 100 virus per 1ml. 000 rabbit infections;

[0032] (3) Pr...

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Abstract

The invention discloses a method for preparing live vaccines of hog cholera and a product thereof. The preparation method comprises the following steps of: (1) culturing porcine passage cell lines; (2) inoculating the porcine passage cell lines with live vaccine production seed viruses of the hog cholera to obtain attenuated vaccine strains of the hog cholera; (3) performing virus multiplication on the attenuated vaccine strains of the hog cholera; (4) measuring the virus titer of multiplication virus suspension by adopting an immunofluorescence method; and (5) adding a freeze-drying protective agent and antibiotics into the virus suspension which is detected to be qualified for vaccine matching and freeze-drying. The preparation method has the advantages of producing the live vaccines of the hog cholera by using the cell lines so as to achieve small quality differences among batches and the characteristics of simple and stable process, easy operation, high yield, low cost, the feasibility and extendibility of industrial production and the like, and measuring the virus titer of the multiplication virus suspension by adopting the immunofluorescence method so as to achieve sensitive, fast, specific and accurate detection, high repeatability and reliable results. The live vaccines of the hog cholera prepared by the method can completely protect pigs from the attacks of violent hog cholera viruses.

Description

Technical field [0001] The invention relates to a production method of a live vaccine for animals, in particular to a preparation method of a swine fever live vaccine and a product obtained by the preparation method, and belongs to the production field of swine fever live vaccine. Background technique [0002] Hog cholera (HC) is a highly contagious infectious disease caused by swine fever virus. The World Organisation for Animal Health (OIE) has included it in the OIE Disease List and is an infectious disease that must be reported by law. In my country, swine fever is one of the major diseases. In the "Category 1, 2 and 3 Animal Diseases List", swine fever is listed as one of the 17 animal diseases in the first category. The outbreak of swine fever has spread to the world and my country. The pig industry caused serious economic losses. [0003] Vaccines are an important means to control swine fever, including inactivated vaccines and attenuated vaccines. Countries are working har...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/187C12N7/08A61P31/14
Inventor 武华
Owner 武华
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