rRT-PCR法在脊髓灰质炎病毒型内鉴定中的应用  

Application of rRT-PCR in the intratypic identification of poliovirus

作　　者：李建雄[1] 熊英[1] 施勇[1] 刘师文[1] 刘晓庆[1] 肖芳[1] 

机构地区：[1]江西省疾病预防控制中心,江西南昌330029

出　　处：《中国卫生检验杂志》2015年第15期2502-2504,2509,共4页Chinese Journal of Health Laboratory Technology

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,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 identify the polioviruses（ PV） using real- time fluorescent quantitative reverse transcription polymerase chain reaction（ rRT- PCR） and evaluate the method. Methods According to rRT- PCR method for polio ITD and VDPV recommended by WHO,the 14 poliovirus isolates from the polio laboratory before 2013 and 15 strains in 2013 were tested for intratypic differentiation（ ITD） and vaccine derived polioviruses（ VDPVs） screening. The results of rRT- PCR were compared with the RNA sequencing detection results on VP1 region of the strains. Results The results of intratypic differentiation of polioviruses using rRT- PCR were consistent with those of VP1 region sequencing except one poliovirus isolate. But the results of VDPV rRT- PCR for poliovirus did not completely consist with those of VP1 region sequencing. 14 strains of type Ⅱ poliovirus were misidentified as vaccine derived polioviruses,and 11 strains of type Ⅲ poliovirus were misidentified as vaccine- like strain. Conclusion rRT- PCR identification method can replace the conventional detection method,but it can not totally replace the sequencing technology for the identification of VDPV.

关 键 词：实时荧光定量逆转录-聚合酶链反应 脊髓灰质炎病毒 型内鉴定 

分 类 号：R512.4[医药卫生—内科学]