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为克服常规反转录ＰＣＲ（ＲＴ－ＰＣＲ）检测肠道病毒的缺点和不足，在实验基础上建立了一步单管ＲＴ－ＰＣＲ检测水中的脊灰病毒，采用热裂解法提取样品中脊灰病毒的总ＲＮＡ，反转录（ｃＤＮＡ）与ＰＣＲ反应在ＰＣＲ仪上一步进行，操作方法简单，中间可能污染的环节少，最大限度地减少了由环境造成的可能污染。设计合成的引物Ｅ１－Ｅ２取自多种肠病毒保守的５′非编码区核酸序列，可用于多种肠道病毒的扩增；Ｐ１－Ｐ２为脊灰病毒３个血清型的共用引物，只扩增脊灰病毒，具有较强的特异性。改进后的ＲＴ－ＰＣＲ的敏感性与常规ＲＴ－ＰＣＲ比有所提高，可检测到细胞培养悬液或水中１０个ＰＦＵ的脊灰病毒。
In order to overcome the shortcomings and deficiencies of routine reverse transcription-polymerase chain reaction (RT-PCR) detection of enteroviruses, one-step single-tube RT-PCR was established to detect poliovirus in water. The poliovirus Of total RNA, reverse transcription (cDNA) and PCR reaction in the PCR instrument step by step, the method of operation is simple, intermediate links may be less pollution, to minimize possible pollution caused by the environment. The designed and synthesized primers E1-E2 are taken from the conserved 5 ’non-coding region nucleic acid sequences of various enteroviruses and can be used for amplification of multiple enteroviruses. P1-P2 are common primer of three serotypes of poliovirus, only Amplification of poliovirus, with strong specificity. The improved RT-PCR sensitivity has been increased compared to conventional RT-PCR, detecting 10 PFU of poliovirus in cell culture suspension or water.