动物医学进展2017，Vol.38Issue(4)：28-34,7.
猪瘟病毒石门株NS3蛋白的原核表达及其抗体检测间接ELISA的建立
Prokaryotic Expression of NS3 Protein in Classical Swine Fever Virus Shimen Strain and Establishment of Indirect ELISA for Antibody Detection
摘要
Abstract
In order to coordinate the new vaccine development,this study established an indirect ELISA for the detection of swine fever virus NS3 protein antibody,to distinguish vaccine immunized pigs and naturally infected pigs(including conventional vaccine vaccination pig)effectively.The total RNA was extracted from PK-15 cells infected with classical swine fever virus(CSFV)Shimen strain,and the CSFV NS3 gene with length of 2 049 bp was obtained by specific PCR amplification.The CSFV NS3 gene was cloned into the prokaryotic expression vector pET-32a(+)with self-immobilized self-cleavage-functional peptide.The expression of NS3 gene was optimized in Escherichia coli Rosetta(DE3).Western blot analysis showed that the recombinant protein NS3 had reactogenicity.The purified recombinant protein NS3 was used as an encapsulated antigen to establish an indirect ELISA for the detection of CSFV NS3 antibody,502 samples were tested on the basis of the antibody test results of Idexx swine fever virus antibody test kit.The result showed that the specificity of this method was 96.9％,the sensitivity was 89.7％,and the total coincidence rate was 95.8％.This research provided the serological diagnosis method for the new vaccine development of classical swine fever.关键词
猪瘟病毒/NS3基因/原核表达/蛋白纯化/间接ELISAKey words
Classical swine fever virus/NS3 protein/prokaryotic expression/protein purification/indirect ELISA分类
农业科技引用本文复制引用
陆欣然,华思红,樊钰莹,白文顺,杜敏,孙永科,杨玉艾..猪瘟病毒石门株NS3蛋白的原核表达及其抗体检测间接ELISA的建立[J].动物医学进展,2017,38(4):28-34,7.基金项目
国家自然科学基金项目(31560704) （31560704）
云南农业大学自然科学基金项目(2016ZR12) （2016ZR12）
