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目的对实时荧光定量逆转录-聚合酶链反应(rRT-PCR)法和微量中和实验2种脊灰病毒的血清型鉴定方法进行评价。方法按世界卫生组织(WHO)《脊髓灰质炎实验室手册》第4版进行病毒分离,用rRT-PCR法和微量中和实验方法对分离到的L20B阳性分离物进行PV的血清型鉴定,用rRT-PCR法进行型内鉴定。结果 rRT-PCR法与微量中和实验对L20B阳性分离物进行脊灰病毒的血清型定型,2种方法检测脊灰病毒阳性率差异无统计学意义,检测NPEV阳性率差异也无统计学意义;rRT-PCR方法检测脊灰病毒的灵敏度为100%,特异度为100%;检测NPEV的灵敏度为63.16%,特异度为100%。结论作为WHO推荐使用的新的型内鉴定方法,rRT-PCR法对于脊灰病毒的血清型定型与常规微量中和实验一致率为100%,该方法方便快捷,可以缩短检测时限;对于NPEV的检测特异度较高,但灵敏度低于微量中和实验。
Objective To evaluate the serological typing of two kinds of polioviruses by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (rRT-PCR) and micro-neutralization assay. Methods The virus was isolated according to the 4th edition of the WHO Polio Laboratory Manual, and the serotypes of PV isolated from L20B positive isolates were identified by rRT-PCR and micro-neutralization test. rRT-PCR method for in-type identification. Results The serotypes of poliovirus serotypes of L20B positive isolates were confirmed by rRT-PCR and micro-neutralization. There was no significant difference between the two methods in detecting the positive rate of poliovirus. There was also no significant difference in the positive rate of detecting NPEV. The sensitivity and specificity of rRT-PCR to detect poliovirus were 100% and 100%, respectively. The sensitivity and specificity of detecting NPEV were 63.16% and 100% respectively. Conclusions As a new intra-type identification method recommended by the WHO, the concordance rate of the rRT-PCR method to the serotype of poliovirus and the conventional micro-neutralization experiment is 100%, which is convenient and fast and can shorten the detection time. For NPEV Detection of higher specificity, but lower than the sensitivity and micro-neutralization experiments.