检测人博卡病毒抗体间接酶联免疫吸附法的建立及临床应用

目的 建立间接酶联免疫吸附法(ELISA)检测人博卡病毒(HBoV)抗体,探讨其临床应用的可行性.方法 以重组表达的HBoV融合蛋白VP2作为包被抗原,建立检测HBoV抗体的间接ELISA,优化该检测方法的最佳条件,观察其精密度、敏感度和特异性,并与荧光定量PCR方法对比检测40份临床样本,同时采用免疫印迹法(Western blot)确认其符合率.结果 所建立的间接ELISA最佳抗原包被浓度为2 mg/L,酶标二抗工作浓度为1∶5000,血清标本最佳稀释度为1∶200.该方法平均批内变异系数为6.87%,平均批间变异系数为4.67%.40份临床样本间接ELISA检测结果与荧光定量PCR及免疫印迹结果一致,符合率100%.结论 利用VP2融合蛋白作为抗原,建立的检测血清HBoV抗体间接ELISA方法特异性强、重复性好,可用于HBoV抗体的定量和定性检测.

Establishment and clinical use of indirect ELISA for detection of human bocavirus (HBoV) antibody

Objective To establish indirect ELISA for detection of HBoV antibody and to evaluate the feasibility of its clinical use. Methods Using the HBoV fusion protein VP2 generated by recombinant expression as envelope antigen, indirect ELISA method for HBoV antibody detection was established. The optimal condition of the test was set, and precision, sensitivity and specificity were examined. The findings of the test were compared with those using quantitative fluorescence PCR in 40 clinical samples, and were validated by Western blot to evaluate the consistency between each other. Results The optimal 4.67%. The findings of this test in 40 clinical samples were 100% consistent with those by quantitative fluorescence PCR and Western blot. Conclusion The new indirect ELISA method for HBoV antibody detection using HBoV fusion protein VP2 as envelope antigen is highly specific, reproducible and useful for quantitative or qualitative detection of HBoV antibody.