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Method for establishing hog cholera lapinized virus labeled vaccine strain and preparing vaccine

A technology of swine fever rabbit and vaccine poison, applied in biochemical equipment and methods, medical preparations containing active ingredients, antiviral agents, etc., can solve problems such as biological safety hazards and unconfirmed immune effects

Active Publication Date: 2011-10-19
CHINA INST OF VETERINARY DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2009 L.G.Holinka et al. (Holinka LG, Fernandez-Sainz I, O'Donnell V, Prarat MV, Gladue DP, Lu Z, Risatti GR, Borca MV Development of a live attenuated antigenic marker classical swine fevervaccine. Virology. 2009: 384( 1): 106-13) Based on the Brescia strain of swine fever virus, a double-labeled vaccine with the deletion of the neutralizing epitope WH303 and the addition of the Flag tag was constructed. The test results showed that the strain had better protection against Brescia strain challenge , but the Brescia strain is virulent and poses a biological safety hazard. In addition, the immune effect of the marked vaccine against other strains has not been confirmed

Method used

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  • Method for establishing hog cholera lapinized virus labeled vaccine strain and preparing vaccine
  • Method for establishing hog cholera lapinized virus labeled vaccine strain and preparing vaccine
  • Method for establishing hog cholera lapinized virus labeled vaccine strain and preparing vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] 1. Vaccine Preparation

[0097] Propagation of venom by using PK15 subcultured cells to produce seedlings

[0098] The transfected cells containing the CGMCC No.4975 strain passed to the 8th to 13th passages were used as seed cells, frozen in liquid nitrogen, and the cells were revived when the virus was propagated. They could be directly cultured or added with 50% negative SK6 cells (, ST and P Any one of the cells such as K15) is co-cultured for about 48-72 hours. After the cells grow into a dense monolayer, the supernatant (virus liquid) is harvested and the cells are passaged at the same time. The harvested virus liquid was frozen and stored below -20°C.

[0099] 2. Inspection of seedling venom:

[0100] (1) Sterility test: according to "Chinese Veterinary Pharmacopoeia" (Chinese Veterinary Pharmacopoeia Committee. People's Republic of China Veterinary Pharmacopoeia 2005 edition three China Agricultural Press, 2006, the present invention is called "Chinese Veterin...

Embodiment 2

[0106] Propagation of venom by using ST subcultured cells to produce seedlings

[0107] The transfected cells containing the CGMCC No.4975 strain passed to the 8th to 13th passages were used as seed cells, frozen in liquid nitrogen, and the cells were revived when the virus was multiplied. They could be directly cultured or co-cultured with 50% negative ST cells, and cultured for 48 After ~72 hours, after the cells grow into a dense monolayer, the supernatant (virus fluid) is harvested, and the cells are passaged at the same time. The harvested virus liquid was frozen and stored below -20°C.

[0108] Other processes of making seedlings are the same as in Example 1.

Embodiment 3

[0110] Propagation of venom by using PK15 subcultured cells to produce seedlings

[0111] The transfected cells containing the CGMCC No.4975 strain passed to the 8th to 13th passages were used as seed cells, frozen in liquid nitrogen, and the cells were revived when the virus was propagated. They could be directly cultured or co-cultured with 50% negative PK15 cells, and cultured for 48 After ~72 hours, after the cells grow into a dense monolayer, the supernatant (virus fluid) is harvested, and the cells are passaged at the same time. The harvested virus liquid was frozen and stored below -20°C.

[0112] Other processes of making seedlings are the same as in Example 1.

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Abstract

The invention relates to a method for establishing a hog cholera lapinized virus labeled vaccine strain and preparing a vaccine. The method comprises the following steps: (1) establishing the overall-length infectious clone of the hog cholera lapinized virus strain (the C strain or the Chinese strain); (2) introducing labeled protein genes; (3) rescuing the hog cholera lapinized labeled vaccine virus; (4) breeding the virus; (5) inspecting a virus solution; (6) preparing a vaccine; and (7) inspecting the finished product of the vaccine, comprising safety inspection and efficacy inspection. The hog cholera lapinized virus labeled vaccine established by the invention maintains the characteristics of good safety, excellent immunogenicity and the like of the hog cholera lapinized virus strain; the virus strain has the advantages of good stability and high cell production virus content, can be used for industrial mass production and can generate a labeled protein specificity antibody aftera hog is immunized to distinguish immunization and natural infection and has important significances on hog cholera prevention, control and purification and emergent immunization.

Description

technical field [0001] The invention relates to the construction of a rabbit-like attenuated labeled vaccine strain of hog fever and a preparation method of the vaccine, belonging to the field of veterinary biological products. Background technique [0002] Classical swine fever virus (CSFV) is an acute, highly lethal, contagious disease of pigs caused by classical swine fever virus (CSFV). According to the 2008 version of the "Terrestrial Animal Diagnostic Test and Vaccine Manual" formulated by OIE, the disease is listed as one of the legally reported animal diseases, and is listed as "Class I Animal Disease" in my country, which is a threat to my country's pig industry. one of the major infectious diseases. [0003] In the early and middle of the 20th century, a large number of researchers developed the Chinese strain of CSF vaccine by different methods (the attenuated strain of CSF vaccine or the Chinese strain of CSFV or the C strain of CSFV), the French Thiverval strain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C12N15/85C12N7/01A61K39/187A61P31/14
Inventor 赵启祖邹兴启朱元源韩焘王琴范学政徐璐宁宜宝刘业兵
Owner CHINA INST OF VETERINARY DRUG CONTROL
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