中国兽医科学2012，Vol.42Issue(3)：258-263,6.
猪捷申病毒8型rVP1-ELISA检测方法的建立
Establishment of an indirect ELISA based on VP1 gene of porcine teschovirus type 8
摘要
Abstract
To establish an indirect ELISA for the detection of antibodies against porcine teschovirus（PTV）,full sequence of gene encoding PTV-8 VP1 protein was cloned into pET-30a vector and the recombinant plasmid was transformed into Rosetta.The recombinant VP1 protein was analyzed by SDS-PAGE and Western-blot.Results indicated that the expressed protein was about 36.2 ku,and was able to specifi-cally react with PTV-8 positive sera.The indirect ELISA was established using the purified recombinant VP1 protein as coating antigen,and the detection limit of the method was 1∶800 of the PTV-8 positive serum dilution.The indirect ELISA established in the present study showed no cross reaction with positive serum of porcine reproductive and respiratory syndrome virus,pseudorabies virus,porcine parvovirus,porcine circovirus type 2,or classical swine fever virus.The coefficient of variation of inter-assay and intra-assay were less than 10%.A total of 633 swine serum samples were tested by the established assay and positive rate was 88.10%,and there was a 94.80% coincidence rate between the indirect ELISA and indirect fluorescence assay based on detection of 58 swine serum samples.关键词
猪捷申病毒/重组VP1蛋白/原核表达/间接ELISAKey words
porcine teschovirus/recombinant VP1 protein/prokaryotic expression/indirect ELISA分类
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胡峰,刘杉杉,蔺文成,王晓玲,张超范,王朝,刘朝霞,胡泉博,崔尚金..猪捷申病毒8型rVP1-ELISA检测方法的建立[J].中国兽医科学,2012,42(3):258-263,6.基金项目
国家自然科学基金资助项目 （）
兽医生物技术国家重点实验室项目 （）
中央级公益性科研院所基本科研业务费专项 （）
