单管中进行反转录－PCR检测肠道病毒RNA

RT PCR OF ENTEROVIRUSE RNA IN A SINGLE REACTION TUBE

采用改进的异硫氰酸胍－酚－氯仿一步抽提法提取肠道病毒ＲＮＡ，通过反复试验，建立了单管一ＲＴ一ＰＣＲ（Ｒｅｖｅｒｓｅｔｒａｎｓｃｒｉｐｔｉｏｎｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ）反应体系。结果表明，此法较常规的分步法简便快速、可靠，为临床检测和流行病学研究提供了较理想的方法。

n order to develope and simplify the procedures of RT一PCR of viral RNA,an improvedmethod,acid guanidinium thiocyanate一phenol一chloroform protocol was used for extraction ofenteroviruse RNA,and RT一PCR was operated in a single reaction tube instead of two tubes.Itis revealed that this protocol is simple,stable,rapid and reliable,is available for detection of en-terovirus RNAs and other viral RNAs. It will be particularly useful for processing large numbersof samples in clinical detection and epidemic study.