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实时荧光定量逆转录-聚合酶链反应在脊髓灰质炎病毒型内鉴定中的应用
Application of rRT-PCR in intratypic identification of poliovirus
【摘要】 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,为进行常规rRT-PCR型内鉴定方法奠定基础。方法采用世界卫生组织(WHO)推荐的rRT-PCR方法,对WHO发放的PV株和实验室分离的PV株进行型内鉴定(ITD)和疫苗衍生PV株(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大部分相符,但有2株Ⅱ型脊髓灰质炎疫苗类似株(SL)PV2-SL被错判为非疫苗类似株(NSL),假阳性率为3.03%;1株Ⅲ型SL被错判为NSL,假阳性率为1.92%。结论在新疆脊髓灰质炎实验室rRT-PCR可以应用于脊髓灰质炎病毒的常规监测及型内鉴定。
【Abstract】 Objective To identify polioviruses using real time fluorescent quantitative reverse transcription poly-merase chain reaction(rRT-PCR) method in Xinjiang Uygur Autonomous Polio Laboratory for the first time, to provide a basis for performing intratypic differentiation by conventional rRT-PCR in polio laboratory. Methods According to rRT-PCR recommended by WHO, poliovirus isolates from the polio laboratory were tested for intratypic differentiation and vaccine-derived polioviruses(VDPVs) screening. Results The results of intratypic differentiation of polioviruses using rRT-PCR were completely consistent with those of VP1 region sequencing. But the results of VDPV rRT-PCR for poliovirus did not completely consist with those of VP1 region sequencing. Two strains of type 2 poliovirus were misidentified as non-Sabin-like viruses with the false positive rate of 3.03% and one trains of type 3 poliovirus were misidentified as non-Sabin-like viruses with the false positive rate of 1.92%. Conclusions The rRT-PCR method can be applied in Xinjiang Polio Laboratory for routine surveillance and intratypic differentiation of poliovirus.
【Key words】 rRT-PCR; Poliovirus; Intratypic differentiation;
- 【文献出处】 疾病预防控制通报 ,Bulletin of Disease Control & Prevention(China) , 编辑部邮箱 ,2015年02期
- 【分类号】R440
- 【下载频次】95