EV71及CA16不同检测方法敏感度及特异性比较

目的比较检测肠道病毒71型(enterovirus 71,EV71)及柯萨奇病毒A组16型(coxsackievirus A16,CA16)不同检测方法、不同生产厂家(A、B、C、D、E)荧光定量PCR检测试剂的敏感度及特异性。方法对EV71及CA16各10个病毒株进行稀释,取原倍、10-3及10-63个浓度,分别采用实时荧光定量PCR法(分别用不同厂家的检测试剂)、RT-PCR法、ELISA法及细胞培养4种方法进行检测,每种检测连续重复3次,然后将其结果进行比较分析。结果 EV71及CA16经不同方法及试剂检测后结果显示,荧光定量PCR方法敏感度最高,依次为细胞培养法、RT-PCR(VP1)法及ELISA法;细胞培养法、RT-PCR(VP1)法及ELISA法的特异性较荧光定量PCR检测方法的特异性高,但荧光定量PCR检测试剂均出现不同程度非特异的假阳性结果;不同的荧光定量PCR检测试剂中,D厂家试剂检测EV71和C厂家试剂检测CA16的敏感度和特异性最高。结论 RT-PCR(VP1)方法更适于检测EV71及CA16,如条件许可则建议联合采用细胞培养检测方法,提高检测结果的敏感度和特异性。

Evaluate the Sensitivity and Specificity of Differentiating Enterovirus 71 and Coxsackievirus A16

Objective To evaluate the sensitivity and specificity of detecting EV71 and CA16 with four different methods and manufacturers(A,B,C,D,E)of the real-time PCR. Methods Ten EV71 strains and ten CA16 strains were diluted to three different concentrations, respectively,the original,10^-3 and 10^-6 times,which were detected simultaneously by the real-time PCR (kits from five manufacturers),RT-PCR,ELISA and Cell culture.Each test repeated three times, and then we compared the results. Results real-time PCR got highest sensitivity, followed by Cell culture, RT-PCR and ELISA. The specificity of cell culture, RT-PCR and ELISA were higher than the real-time PCR, but there were non-specificity and false positive result by the method of the real-time PCR. EV71 from D manufacturer achieved the highest sensitivity and specificity. CA16 from C manufacturer achieved the highest sensitivity and specificity. Conclusion RT-PCR (VP1) is more suitable for detecting EV71 and CA16. It suggests that combining cell culture will improve sensitivity and specificity if the condition allows.