RT-PCR-RFLP方法用于脊髓灰质炎病毒型内鉴定  

Intratypic Differentiation of Poliovirus by Using RT-PCR-RFLP

作　　者：田宏[1] 刘杨[1] 高志刚[1] 万丽霞[1] 

机构地区：[1]天津市卫生防病中心,300011

出　　处：《天津医药》2004年第8期485-486,F005,共3页Tianjin Medical Journal

摘　　要：目的 :采用逆转录 -聚合酶链反应 -限制性酶切片段长度多态性分析 (RT_PCR_RFLP)方法对脊髓灰质炎(脊灰)病毒 (PV)进行型内鉴定。方法 :采用RT_PCR方法对PV的VP1区进行扩增 ,PCR产物分别用DdeⅠ、HpaⅡ、HaeⅢ3种限制性内切酶做RFLP分析 ,并与PVSabinⅠ、Ⅱ、Ⅲ3个型别疫苗株相对照判定其是否为疫苗相关株。结果 :对天津市2001年—2002年分离到的9株PV阳性株进行RT_PCR_RFLP试验 ,全部为疫苗相关株。结论 :该方法应用于PV检测 ,可以尽早发现脊灰野病毒、疫苗衍生株及疫苗重组株等 ,便于尽快采取措施阻断其传播。Objective:To use a method of reverse transcriptase-polymerase chain reaction restriction fragˉment length polymorphism(RT_PCR_RFLP)for intratypic differentiation of poliovirus.Methods:VP1fragˉment coding regions of polioviruses were amplified with RT_PCR methods.The amplified products were diˉgested with DdeⅠ,HpaⅡ,and HaeⅢrestriction endonucleases for the analysis of RFLP.Compared the reˉsults of RFLP to Sabin_like viruses,it was decided if the testing strains were Sabin_like viruses.Results:Nine poliovirus isolates from Tianjin during2001to2002were tested with RT_PCR_RFLP.All of them were Sabin_like viruses.Conclusion:Application of the method is available of finding wild poliovirus,vaccine_derived poliovirus and recombinant strain,and preventing them from spreading.

关 键 词：RT—PCR—RFLP法 脊髓灰质炎病毒 疫苗衍生株 疫苗重组株 

分 类 号：R446.5[医药卫生—诊断学]