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目的在河北省脊髓灰质炎(脊灰)实验室首次应用实时荧光定量逆转录-聚合酶链反应(real time fluorescent quantitative reverse transcription-polymerase chain reaction,rRT-PCR)对脊灰病毒(PV)进行鉴定,并对该方法进行评估,为进行常规rRT-PCR型内鉴定方法做准备。方法采用世界卫生组织(WHO)推荐的rRT-PCR方法,对河北省既往分离的PV和WHO发放的PV株进行型内鉴定(intratypic differentiation,ITD)和疫苗衍生PV(vaccine-derived PV,VDPV)筛选。结果 ITD rRT-PCR的实验结果与毒株的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能完全相符,有2株Ⅱ型脊灰病毒被错判为NSL,假阳性率为6.9%(2/29)。结论 Real time PCR脊灰型内鉴定方法可以在河北省脊灰实验室用于脊灰病毒的常规监测。
Objective To identify poliovirus (PV) for the first time by real time fluorescent quantitative reverse transcription-polymerase chain reaction (rRT-PCR) in the polio laboratory of poliomyelitis in Hebei Province , And the method was evaluated for preparation for routine rRT-PCR type identification. Methods According to the rRT-PCR method recommended by the World Health Organization (WHO), intratypic differentiation (ITD) and vaccine-derived PV (VDPV) filter. Results The results of ITR rRT-PCR were in good agreement with those of the VP1 coding region of the strain. The results of VDPV rRT-PCR and the results of VP1 coding region were not completely consistent. Two strains of type Ⅱ poliovirus were mistakenly identified as NSL, the false positive rate was 6.9% (2/29). Conclusion Real time PCR can be used to detect poliovirus in poliovirus laboratory in Hebei Province.