ELISA法检测人微小B19抗体及其临床应用评价

Clinical value of measuring human parvovirus B19 antibody by ELISA

目的建立检测人微小病毒B19的间接酶联免疫吸附(ELISA)法,评价其临床应用价值。方法采用原保存的XA-B19 VP1独特区蛋白包被ELISA板,优化该方法检测B19抗体的最佳实验条件。与聚合酶链反应(PCR)、parvovirus B19 ELISA方法进行比较,评价其一致性。结果最佳包被量为25 ng/孔,标本血清最佳稀释倍数为1∶200。建立的间接ELISA检测体系与腺病毒、呼吸道合胞病毒、流感病毒、副流感病毒、疱疹病毒抗体阳性血清无交叉反应。其检测B19 IgM敏感性为88.37%,特异性为96.15%,与PCR方法一致性好(kappa值>0.75,P>0.05);与parvovirus B19 IgM ELISA方法符合性好,符合率为96.8%。与parvovirus B19IgG ELISA方法比较kappa值>0.75,P>0.05,两种检测方法一致性好。结论建立的间接ELISA检测B19抗体的方法具有灵敏度高、特异性强、经济、快速、方便等优点,适合于流行病学调查和临床标本检测。

Objectives To establish indirect enzyme linked immunosorbent assay （ELISA） method for detection of human parvovirus B19 antibody and evaluate its importance for clinical application. Methods Specific B19 antibodies were detected with indirect ELISA. The method was further optimized by coating ELISA plate with XA-B19 VP1 unique recombinant protein. Comparison among this indirect ELISA method, polymerase chain reaction （PCR） and parvovirus B19 IgG, IgM ELISA kit （Germany） was done, and the consistency among these three methods was assessed as well. Results The optimal concentration of coated antigen VP1 unique recombinant protein was 25 ng per well and the optimal serum dilution was 1：200. This indirect ELISA system has no cross-reaction with the serum antibodies of adenovirus, respiratory syncitial virus, influenza virus, parainfluenza virus, and human herpes virus. The sensitivity was 88.37% and the specificity was 96.15% when it was used to detect B19 IgM. The concordance between the indirect ELISA method and PCR was good （kappa 〉 0.75, P 〉 0.05）. The concordance between the indirect ELISA method and parvovirus B19 IgM ELISA kit was also good. When it was used to detect B19 IgG, the concordance between the indirect ELISA method and parvovirus B19 IgG ELISA kit was good （kappa 〉 0.75, P 〉 0.05）. Conclusions Indirect ELISA is sensitive, specific, economical, quick and convenient for detection of antibodyagainst parvovirus B19 and it is suitable for clinical use.