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目的应用实时荧光定量逆转录-聚合酶链反应(rRT-PCR)对脊髓灰质炎病毒(PV)进行型内鉴定,并对该方法进行评估。方法采用世界卫生组织(WHO)推荐的脊髓灰质炎ITD和VDPV rRT-PCR方法对江西省既往分离的14株脊髓灰质炎毒株和2013年分离的15株脊髓灰质炎毒株进行ITD和VDPVs筛选,并将检测结果与毒株的VP1编码区核苷酸序列测定结果进行比较分析。结果 ITD rRT-PCR的实验结果除1株毒株漏检外,其余与毒株的VP1编码区序列测定结果完全相符,VDPV rRT-PCR的结果与VP1编码区序列测定结果不完全相符,共有14株Ⅱ型脊髓灰质炎病毒疫苗类似株被错判为VDPV株,11株Ⅲ型VDPV株错判为疫苗类似株。结论对脊髓灰质炎型内进行rRTPCR鉴定的方法可以替代中和试验的常规检测方法,但不能完全取代测序技术用于脊髓灰质炎VDPV的鉴定。
Objective To evaluate the in vivo characterization of poliovirus (PV) by real-time quantitative reverse transcription-polymerase chain reaction (rRT-PCR). Methods 14 poliomyelitis isolates previously isolated in Jiangxi Province and 15 poliomyelitis isolates isolated in 2013 were screened for ITD and VDPVs using the polio ITD and VDPV rRT-PCR methods recommended by the World Health Organization (WHO) , And the test results were compared with the results of nucleotide sequence determination of the VP1 coding region of the strain. Results The result of ITR rRT-PCR was identical to that of the VP1 coding region except for one strain except for one strain, and the results of VDPV rRT-PCR did not exactly match the results of VP1 coding region The strain of type II poliovirus vaccine was mistakenly identified as VDPV strain, and 11 strains of type III VDPV strain were mistakenly identified as vaccine-like strains. Conclusion The method of rRTPCR identification in poliovirus can replace the conventional detection method of neutralization test, but it can not completely replace the sequencing technology for the identification of polio VDPV.