快速诊断人细小病毒B19感染方法学比较研究

Comparison of different diagnostic methods for rapid detection of human parvovirus B19 infection

目的比较本实验室自行构建的ELISA法与德国ELISA试剂盒方法及PCR法快速诊断人细小病毒B19感染的临床应用价值。方法血液标本215份,分别采用B19病毒VP1独特区(VP1u)蛋白包被ELISA板建立B19病毒抗体检测方法(自行构建ELISA法),德国ELISA试剂盒方法检测标本B19病毒IgM与IgG,并采用PCR法检测B19病毒DNA,分析自行构建ELISA法检测B19病毒的敏感性、特异性及准确度。结果自行构建的B19病毒ELISA体系最佳抗原包被量为25ng/孔,最佳标本血清稀释倍数为1∶200;以德国B19病毒ELISA试剂盒为对照,自行构建ELISA法检测B19病毒IgM的敏感性、特异性及准确度分别为79.07%,87.18%,94.89%,检测B19病毒IgG的敏感性、特异性及准确度分别为88.46%,90.79%,93.53%;以PCR法检测B19病毒DNA为标准,自行构建ELISA法检测B19病毒IgG的敏感性、特异性及准确度分别为90.32%,77.77%,95.71%。结论本实验室自行构建的ELISA检测方法与德国B19病毒抗体检测ELISA试剂盒法及PCR法检测B19病毒DNA有较好一致性,可作为B19病毒感染常规诊断方法。

Objective To compare and evaluate the ELISA method constructed by our laboratory with human parvovirus B19 （B19）, German ELISA Kit and PCR technique for detecting B19 infection. Methods Our own ELISA method was modified by being coated with B19 VP1 unique （VPlu） recombinant protein. IgM and IgG were detected with the modified ELISA method and German B19 ELISA Kit in 215 blood samples. B19 DNA was detected with PCR technique. The sensibility, specificity and accuracy were analyzed and compared. Results The optimal quantity of coated VPlu protein was 25 ng per well, and the optimal dilution of sample was 1 ： 200. The sensibility, specificity and accuracy were 79.07%, 87.18% and 94.89% for B19 IgM, and 88.46%, 90.79% and 93.53% for B19 IgG compared with German B19 ELISA Kit, and were 90.32%, 77.77% and 95.71% for B19 IgG compared with PCR technique. Conclusion Our own modified ELISA method has good concordance with German B19 ELISA Kit and PCR technique, and can be used as a routine clinical diagnosis method for B19 infection.