中国实用儿科杂志2009，Vol.24Issue(10)：769-771,3.
人细小病毒B19 VP1蛋白表达及检测特异性抗体临床应用研究
Expression of human parvovirus B19 VP1 protein and its clinical application to detection of specific antibody
摘要
Abstract
Objective To establish enzyme-linked immunosorbent assay(ELISA)method for detection of human parvo-virus B19 antigen and compare it with parvovirus B19 IgG 、IgM ELISA Kit (Germany) to determine its sensitivity,speci-ficity and accuracy. Methods Constructed VP1-PQE30 was transformed into E.coli M15 and was analysed by se-quencing. The E.coli M15 with VP1-PQE30 were fermented and induced by IPTG to express confluenced VP1 protein . SDS-PAGE and Western blot were used to evaluate the expressed protein. The expressed confluenced VP1 protein was purified by AKTA explorer 100 system. After determining the optimal concentration of coated antigen and the optimal di-lution of blood serum, establish enzyme-linked immunosorbent assay (ELISA) method for detection of human parvovirus B19 antigen and evaluate its value for clinical application. Results This ELISA system had no cross-reaction with the serum antibodies of adenovirus, respiratory syncytial virus, influenza virus, parainfluenza virus, and human herpes virus. The sensitivity of detection of IgM and IgG was 87.50% and 89.28%, the specificity 90.34%and 90.90% and the accura-cy 97.01% and 95.54%. There was a good concordance between the two methods. Conclusions It is sensitive, specific and precise to use domestic-made B19 VP1 unique recombinant protein to detect human parvovirus B19 antigen. It is quick and economical, so it is worth investigating deeply and spreading .关键词
人细小病毒B19/VP1独特区蛋白/ELISA/原核表达Key words
human parvovirus B19/ VP1 unique protein/ enzyme-linked immunos- orbent assay/ prokaryotic express分类
医药卫生引用本文复制引用
许东亮,张国成,聂晓晶,李志宏,孙新,张学红..人细小病毒B19 VP1蛋白表达及检测特异性抗体临床应用研究[J].中国实用儿科杂志,2009,24(10):769-771,3.基金项目
国家自然科学基金资助项目[30571948] （）
