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Preparation method and product of swine fever live vaccine

A swine fever live vaccine and vaccine strain technology, which can be applied to medical preparations containing active ingredients, pharmaceutical formulations, inactivation/attenuation, etc. Accurate quantification, long test period and other problems, to achieve the effect of swine fever virulent attack protection, simple and stable production process, and small quality difference

Active Publication Date: 2011-12-28
华威特(江苏)生物制药有限公司
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  • 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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  • Preparation method and product of swine fever live vaccine
  • Preparation method and product of swine fever live vaccine
  • Preparation method and product of swine fever live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Screening and identification of pig-derived cell lines

[0027] Inoculate different pig-derived cell lines with attenuated classical swine fever rabbit virus, culture at 37°C for 4-5 days, harvest the virus, pass the harvested virus, and use immunofluorescence method to detect each generation of virus inoculated with different pig-derived cell lines , the test results show that the attenuated CSF rabbit can be tested in PT cell lines, MPK cell lines, SK6 cell lines, PK 2a cell lines, CPK cell lines, RKC cell lines, MDBK cell lines, MDCK cell lines, CRFK cell lines, Vero / Proliferate on slam cell line, Vero-E6 cell line, Marc-145 cell line, BHK cell line, BGM cell line or DK cell line. The present invention uses cell lines (PT, MPK, SK6, PK 2a, CPK, RKC, MDBK, MDCK, CRFK, Vero / slam, Vero-E6, Marc-145, BHK, BGM or DK) to cultivate attenuated swine fever rabbits , harvest the cell virus liquid, use the immunofluorescence method to measure the toxicity of the pro...

Embodiment 2

[0028] Embodiment 2 produces the method for swine fever live vaccine with cell line

[0029] (1) Passage and culture of cells for seedling production: PT cell lines were digested and passaged with trypsin-EDTA cell dispersion, and passaged at a ratio of 1 to 5. Continue culturing at 37°C with cell growth medium, and when a good monolayer is formed, it is used for continued passage or virus inoculation;

[0030](2) Propagation of cytotoxic species: use cell maintenance solution to make 0.3% virus suspension from fresh spleen venom, inoculate a single layer of well-grown PT cell line, and place at 37°C to continue culturing. Harvest the cell culture venom every 5 days as the virus species for production; identification of the cytotoxic species: the identification of the cytotoxic species fully complies with the standard of the attenuated strain of classical swine fever, which is safe for pigs and has no side effects. Each 1ml of the cytotoxic species contains >100,000 viruses R...

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Abstract

The invention discloses a preparation method and product of a swine fever live vaccine. The preparation method comprises the following steps: (1) culturing a swine-derived continuous cell line; (2) inoculating the swine-derived continuous cell line into a seed virus for producing the swine fever live vaccine, thereby obtaining a swine fever weak-virus vaccine strain; (3) carrying out virus multiplication on the swine fever weak-virus vaccine strain; (4) determining the virus value of the multiplied virus suspension by an immunofluorescence method; and (5) adding a freeze-drying protective agent and antibiotics into the qualified virus suspension to carry out vaccine preparation and freeze-drying. Since the swine fever live vaccine is prepared from the cell line, the difference of quality among different batches is small, and the invention has the characteristics of simple and stable technique, high yield, low cost and the like, is easy to operate, and has feasibility and amplification of industrialized mass production. In addition, the immunofluorescence method, which has the advantages of high detection sensitivity, high speed, high specificity, high accuracy, high repetitiveness and reliable result, is used for determining the virus value of the multiplied virus suspension. The swine fever live vaccine disclosed by the invention can 100% protect the attack of swine fever strong virus.

Description

technical field [0001] The invention relates to a production method of a veterinary live vaccine, in particular to a preparation method of a live swine fever vaccine and a product obtained by the preparation method, and belongs to the field of production of a live swine fever vaccine. Background technique [0002] Hog cholera (HC) is a highly contagious disease caused by classical swine fever virus. The World Organization 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 epidemic diseases. It is listed as one of the 17 kinds of animal diseases in the "List of Types I, II, and III Animal Epidemic Diseases". The pig industry has caused serious economic losses. [0003] Vaccines are an important means of controlling swine fever, including inactivated vaccines and attenuated vaccines. Countries around the world are working hard to develop safe and effective va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/187C12N7/08A61P31/14
Inventor 武华夏铭崎张淑琴
Owner 华威特(江苏)生物制药有限公司
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