实时荧光定量逆转录-聚合酶链反应脊灰型内鉴定新方法在脊灰实验室的应用  

Application of rRT-PCR Method in Intratypic Identification of Poliovirus in a Polio Laboratory

作　　者：陈玫[1] 赵娜[1] 郭玉[1] 崔志强[1] 张振国[1] 

机构地区：[1]河北省疾病预防控制中心免疫规划管理所,河北石家庄050021

出　　处：《实用预防医学》2013年第11期1389-1392,共4页Practical Preventive Medicine

摘　　要：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 the polioviruses firstly using real time fluorescent quantitative reverse transcription polymerase chain reaction （rRT-PCR） method in Hebei Provincial Polio Laboratory,to evaluate the assay and provide a basis for performing intratypic differentiation by conventional rRT-PCR in the 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 polio-virus 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 6.9 ％ （2/29）.Conclusions The rRT-PCR method can be used for routine surveillance of poliovirus in Hebei Provincial Polio Laboratory.

关 键 词：实时荧光定量逆转录-聚合酶链反应 疫苗衍生脊灰病毒 疫苗类似株 

分 类 号：R512.4[医药卫生—内科学]