脊髓灰质炎病毒三类检测方法的灵敏度比较

Sensitivity Comparison of Several Detection Assays for Poliovirus

目的对三种脊髓灰质炎病毒检测方法的灵敏度进行比较。方法将I、Ⅱ和Ⅲ型脊灰病毒标准株（TR株），从1×10^1倍系列稀释至1×10^7倍。选用三类方法对制备的脊灰病毒稀释液系列进行检测，比较不同检测方法的灵敏度，三类方法分别为：RD和L20B细胞分离脊灰病毒、肠道病毒通用引物EVl／EV2和脊灰病毒特异性引物UC11／UGl分别进行RT—PCR检测脊灰病毒核酸、三种商业性肠道病毒Realtime RT—PCR检测试剂盒检测脊灰病毒核酸。结果RD和L20B细胞分离脊灰病毒的检测限可达脊灰病毒标准株的lO。倍稀释度；基于肠道病毒通用引物（EVl／EV2）和脊灰病毒通用引物（UC11／UGl）的两种普通RT—PCR检测限分别为10^3、10^2倍稀释度；达安、金豪、基因格三种商业性实时荧光RT—PCR试剂盒的检测限分别为：10^1、10^3、10^4倍稀释度。结论RD和L20B两种细胞分离脊灰病毒的灵敏度并不亚于现有的核酸检测方法。目前部分商业性的肠道病毒通用型核酸检测试剂盒主要针对手足口设计，对脊灰病毒的检测灵敏度反而不够，应慎重选用。

Objectiveto To compare the sensitivity of several assays for poliovirus detection. Methods Three serotypes of poliovirus standards（ Ⅰ ,Ⅱ andⅢ ） were serially deluted from 1 × 10^1 fold to 1 × 10^7 fold respectively. These dilution series were tested by three kinds of methods, including virus isolation assay using RD and L20B cells, RT-PCR assays using enterovirus group specific primer sets （EV1/ EV2 ） and poliovirus group specific primer sets （UCI 1/UG1 ）, and real time RT-PCR assays including three available commercial real time RT-PCR kits. Results The detection limit of the virus isolation method is 1 × 10^4 fold. The sensitivities of EV1/EV2 based and UCll/UG1 based RT-PCRs were 1 × 10^3 fold and 1 × 10^2 fold respectively. The detection limit of three real time PCR kits （Da An, King Hawk and Genegle） was 1 × 10^1 fold, 1 × 10^3 fold and 1 × 10^4 fold. Conclusion the sensivivity of virus isolation method is not less than that of any nucleid acid based methods. Some commercial real time PCR kits were developed initially for HFMD detection and were not suit for polio virus detection.