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逆转录-聚合酶链反应(RT-PCR)技术同时检测水中多种肠道病毒

张崇淼 刘永军 王晓昌 薛小平

张崇淼, 刘永军, 王晓昌, 薛小平. 逆转录-聚合酶链反应(RT-PCR)技术同时检测水中多种肠道病毒[J]. 环境科学研究, 2007, 20(3): 137-141.
引用本文: 张崇淼, 刘永军, 王晓昌, 薛小平. 逆转录-聚合酶链反应(RT-PCR)技术同时检测水中多种肠道病毒[J]. 环境科学研究, 2007, 20(3): 137-141.
ZHANG Chong-miao, LIU Yong-jun, WANG Xiao-chang, XUE Xiao-ping. Simultaneous Detection of Enteroviruses in Water Samples by RT-PCR[J]. Research of Environmental Sciences, 2007, 20(3): 137-141.
Citation: ZHANG Chong-miao, LIU Yong-jun, WANG Xiao-chang, XUE Xiao-ping. Simultaneous Detection of Enteroviruses in Water Samples by RT-PCR[J]. Research of Environmental Sciences, 2007, 20(3): 137-141.

逆转录-聚合酶链反应(RT-PCR)技术同时检测水中多种肠道病毒

基金项目: 国家自然科学基金资助项目(50478048);国家自然科学基金重大国际合作项目(50621140001)

Simultaneous Detection of Enteroviruses in Water Samples by RT-PCR

  • 摘要: 建立了一种利用通用引物RT-PCR技术检测水中肠道病毒的方法.利用脊髓灰质炎病毒1~3型,柯萨奇病毒B3型作为参考病毒株,根据肠道病毒RNA5′非编码区中具有高度同源性的序列来设计通用引物.比较了M-MLV酶和AMV酶的逆转录效果,AMV酶能够成功地从地表水和生活污水中逆转录病毒RNA,更适于实际应用.对比研究了PCR过程中的退火温度,c(Mg2+)等因素对RT-PCR检测结果的影响,选择退火温度55 ℃,c(Mg2+)为2 mmol/L的反应条件,优化了RT-PCR检测方法.通过检测水样中接种的连续稀释的病毒,确定了该检测方法的灵敏度为38 CCID50.考察人工污染的地表水、污水、二级处理出水样品发现,检测灵敏度基本一致.该方法可应用在实际环境的肠道病毒检测中.

     

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  • 刊出日期:  2007-06-25

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