中文摘要：

目的建立能同时检测水环境中肠道病毒（Enterovirus，EVs）、诺如病毒GⅡ（Norovorus，Nov GⅡ）、轮状病毒（Rotavirus，RVs）、腺病毒（Adenovirus，AdVs）的多重RT—PCR检测方法。方法根据GenBank收录的肠道食源性病毒核酸序列，利用软件DNAMAN设计肠道病毒通用引物以及NovGⅡ、RVs、AdVs的特异性引物；将这4对引物置于同一体系中进行多重RT—PCR，利用各种肠道食源性病毒的核酸模板检测其特异性并通过不同滴度[100-107pfu（copies）／ml]病毒测试其灵敏度；利用加标实验验证该方法的准确性；通过对5份河水或某水厂出水大体积水样（50L）进行病毒富集与检测，以确定该方法的实用性，同时，采用传统细胞培养法验证其结果。结果该方法对目的病毒均获得了理想的电泳条带，而对其他病毒无非特异扩增；对EVs、AdVs的最低检出滴度均为10pfu／ml，NovGⅡ—4为10copies／ml，对RVs的最低检出滴度为102pfu／ml；加标实验检测结果准确率为100％；对5份河水或某水厂出水大体积水样进行病毒富集与检测，对以上各种病毒均有检出且与细胞培养结果一致。结论该方法不仅特异性强、灵敏度高、简便快捷，而且准确率高，实际应用性强，适用于水中肠道食源性病毒的快速检测。

英文摘要：

Objective To develop a multiplex reverse transcription polymerase chain reaction method （multiplex RT-PCR） for the detection of enterovirus （EV）, rotavirus （RV） Norovirus G Ⅱ and adenovirus （AdVs） in water environment. Methods According to gene of intestinal viruses included in GenBank, DNAMAN was used to design primers for EV, G Ⅱ --4, RVs, AdVs. Then, these four pairs of primers were put in the same PCR system to test specificity with different viral nucleic acid templates[100-107 pfu（copies）/ml], sensitivity with different viral titer and accuracy with standard experiment. At the same time, this method was applied to concentrate and detect the viruses in seven samples from five rivers and two finished water samples to verify the practicability, while validated the results with the traditional cell culture method. Results The multiplex RT-PCR had good specificity, getting an ideal stripe for the testing virus. The sensitivities for EV, AdVs were 10 pfu/ml, Nov G Ⅱ -4 was 10 copies/ml, and for RVs is 102 pfu/ml; Accuracy of standard experiment was 100%. This method was also used to detect viruses in concentrate from 50 L water sample and all viruses above were detected successfully and the result was consistent with that of cell culture method. Conclusion The multiplex RT-PCR is sensitive,simple, rapid, accurate and applicable to the detection of intestinal viruses in water.