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Safe carrier for seed viruses of inactivated vaccine for foot-and-mouth disease and application thereof

A technology for foot-and-mouth disease virus and foot-and-mouth disease, which is applied in inactivation/attenuation, virus/phage, introduction of foreign genetic material using vectors, etc., can solve the problems of incomplete vaccine inactivation, incomplete virus aggregation and inactivation, etc. The effect of force reduction and high safety

Active Publication Date: 2013-04-03
CHINA INST OF VETERINARY DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Formaldehyde is the inactivator used in the foot-and-mouth disease inactivated vaccine initially, but because formaldehyde inactivates the coagulation protein, it is easy to cause a large number of viruses to accumulate and inactivate incompletely, and it was later replaced by BEI. Incompletely related, the EU stopped using vaccines in 1990
Developed countries such as Europe and the United States have established vaccine antigen banks for foot-and-mouth disease to prepare emergency vaccines, but they clearly stipulate that inactivated antigens cannot be produced in their own countries. The preserved antigens are all purchased and stored after production and inactivation by manufacturers in Africa and South America.

Method used

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  • Safe carrier for seed viruses of inactivated vaccine for foot-and-mouth disease and application thereof
  • Safe carrier for seed viruses of inactivated vaccine for foot-and-mouth disease and application thereof
  • Safe carrier for seed viruses of inactivated vaccine for foot-and-mouth disease and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] ——Construction of type A foot-and-mouth disease chimeric virus and vaccine production

[0106] One, the construction of type A foot-and-mouth disease chimeric virus

[0107] 1. Construction of infectious clones of type A foot-and-mouth disease chimeric virus: A / HuBWH / CHA / 2009 strain P1 of type A (primers No.12, No.13) (Shanghai Yingjun Biotechnology Co., Ltd.) was obtained by PCR amplification Genes (including VP2, VP3, VP1), while introducing restriction sites Ssp Ⅰ and SgrA Ⅰ (SEQ ID NO: 18) at both ends, after cloning and sequencing, positive colonies were screened, bacterial culture, plasmid extraction, and Ssp Ⅰ / SgrA Ⅰ double Restriction digestion and ligation into p43-CHA90-LL-Vector to obtain type A chimeric virus cDNA infectious clones.

[0108] 2. In vitro transcription of chimeric virus cDNA infectious clone: ​​linearize type A chimeric virus cDNA infectious clone plasmid, and transcribe into RNA by in vitro transcription kit.

[0109] 3. Virus rescue: Type ...

Embodiment 2

[0136] ——Construction and vaccine production of O-type foot-and-mouth disease Cathay topological chimeric virus

[0137] 1. Construction of O-type foot-and-mouth disease Cathay topological chimeric virus

[0138] 1. Construction of infectious clones of O-type foot-and-mouth disease Cathay topological chimeric virus: Obtain Cathay topological type new swine virus Ⅱ pedigree O / The P1 gene (including VP2, VP3, VP1) of the GX / 09-7 strain, at the same time, introduced restriction sites Ssp Ⅰ and SgrA Ⅰ (SEQ ID NO: 21) at both ends, cloned and sequenced, screened positive colonies, cultured bacteria, and extracted plasmids , was ligated into p43-CHA90-LL-Vector by Ssp Ⅰ / SgrA Ⅰ double enzyme digestion, and the cDNA infectious clone of O-type foot-and-mouth disease Cathay topological chimeric virus was obtained.

[0139] 2. In vitro transcription of chimeric virus cDNA infectious clone: ​​linearize the cDNA infectious clone plasmid of O-type foot-and-mouth disease Cathay topological...

Embodiment 3

[0165] ——Construction of O-type foot-and-mouth disease ME-SA topological chimeric virus and vaccine production

[0166] 1. Construction of Chimeric Viruses of Type O Foot-and-Mouth Disease ME-SA Topological Pan-Asian Strains

[0167] 1. Construction of infectious clones of O-type FMD ME-SA topological chimeric virus: ME-SA topological pan-sublineage was obtained by PCR amplification (No.16, No.17) (Shanghai Yingjun Biotechnology Co., Ltd.) The P1 gene (including VP2, VP3, VP1) of the O / HB / 07-4 strain, at the same time, introduced restriction sites Ssp Ⅰ and SgrA Ⅰ (SEQ ID NO: 20) at both ends, cloned and sequenced, screened positive colonies, cultured the bacteria, The plasmid was extracted and ligated into p43-CHA90-LL-Vector with Ssp Ⅰ / SgrA Ⅰ double enzyme digestion to obtain cDNA infectious clones of O-type foot-and-mouth disease ME-SA topological pan-sublineage chimeric virus.

[0168] 2. In vitro transcription of the chimeric virus cDNA infectious clone: ​​linearize the ...

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Abstract

This invention relates to a safe carrier for the seed viruses of inactivated vaccine for the foot-and-mouth disease and the application thereof. Different serum-type seed viruses for producing the inactivated vaccines for the foot-and-mouth disease safely are constructed on the basis of the carrier. The constructed embedded viruses retain the advantage of good adaptability of the classic vaccine virus strain cell, overcome the defect of low matching performance of the antigen through the structural protein inserted into the epidemic virus strain and can save virus quickly to produce the inactivated vaccine for foot-and-mouth disease. The A or Asia I or O type different virus-strain embedded viruses which are constructed through the carrier are less virulent and have good bio-security. Because the viruses are short of the protein L, the virulence of the viruses can not become stronger under the natural condition, and the pathogenicity of the viruses on the neonatal rat, pig or cow is reduced greatly or the viruses are not pathogenic. The carrier is highly safe, can not cause the foot-and-mouth disease and is highly practical.

Description

technical field [0001] The invention relates to a safe seed virus carrier of an inactivated foot-and-mouth disease vaccine and its application, that is, the safe production seed virus of an inactivated foot-and-mouth disease vaccine of different serotypes is constructed on the basis of the carrier. It belongs to the field of veterinary biological products. Background technique [0002] Foot-and-mouth disease is an acute, febrile, highly contagious viral disease of artiodactyls caused by foot-and-mouth disease virus. Except for some islands, the disease was widely spread all over the world, causing huge economic losses to the world's animal husbandry. Countries around the world have invested heavily in controlling and eradicating foot-and-mouth disease. As of May 2007, OIE announced that 59 countries are non-immune. Among the FMD-free countries, parts of 9 countries are non-immune FMD-free areas, and 7 countries or regions are immune to FMD-free countries. Foot-and-mouth di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N1/21C12N7/04C12N7/00A61K39/135A61P31/14C12R1/19
Inventor 赵启祖邹兴启朱元源徐璐范学政王琴沈青春王芳宁宜宝
Owner CHINA INST OF VETERINARY DRUG CONTROL
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