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首页> 外文期刊>Applied Microbiology >Development and Evaluation of EPA Method 1615 for Detection of Enterovirus and Norovirus in Water
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Development and Evaluation of EPA Method 1615 for Detection of Enterovirus and Norovirus in Water

机译：用于检测水中肠病毒和诺如病毒的EPA方法1615的开发和评估

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The U.S. EPA developed a sample concentration and preparation assay in conjunction with the total culturable virus assay for concentrating and measuring culturable viruses in source and drinking waters as part of the Information Collection Rule (ICR) promulgated in 1996. In an effort to improve upon this method, the U.S. EPA recently developed Method 1615: Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Method 1615 uses a culturable virus assay with reduced equipment and labor costs compared to the costs associated with the ICR virus method and introduces a new molecular assay for the detection of enteroviruses and noroviruses by reverse transcription-quantitative PCR. In this study, we describe the optimization of several new components of the molecular assay and examine virus recovery from ground, reagent-grade, and surface water samples seeded with poliovirus type 3 and murine norovirus. For the culturable virus and molecular assays, mean poliovirus recovery using the complete method was 58% and 20% in groundwater samples, 122% and 39% using low-titer spikes in reagent-grade water, 42% and 48% using high-titer spikes in reagent-grade water, and 11% and 10% in surface water with high turbidity, respectively. Murine norovirus recovery by the molecular assay was 30% in groundwater samples, less than 8% in both low- and high-titer spikes in reagent-grade water, and 6% in surface water with high turbidity. This study demonstrates the effectiveness of Method 1615 for use with groundwater samples and highlights the need for further research into its effectiveness with surface water.
机译：美国EPA作为1996年颁布的《信息收集规则》（ICR）的一部分，开发了一种样品浓度和制备分析方法以及总可培养病毒分析法，用于浓缩和测量源水和饮用水中的可培养病毒。作为一种方法，美国EPA最近开发了方法1615：通过培养和RT-qPCR测量水中肠道病毒和诺如病毒的发生率。与使用ICR病毒方法的成本相比，方法1615使用了可培养的病毒测定法，其设备和人工成本降低了，并引入了一种新的分子测定法，用于通过逆转录定量PCR检测肠病毒和诺如病毒。在这项研究中，我们描述了分子测定法几个新组成部分的优化，并检查了从接种了3型脊髓灰质炎病毒和鼠诺如病毒的地面，试剂级和地表水样品中回收的病毒。对于可培养的病毒和分子分析，使用完整方法的脊髓灰质炎病毒的平均回收率在地下水样品中为58％和20％，在试剂级水中使用低滴度加标时分别为122％和39％，使用高滴定度时为42％和48％试剂级水的峰值增加，高浊度的地表水中的峰值分别达到11％和10％。通过分子分析，鼠诺如病毒的回收率在地下水样品中为30％，在试剂级水的低滴度和高滴度加标中均不到8％，在高浊度的地表水中仅为6％。这项研究证明了方法1615用于地下水样品的有效性，并强调了需要进一步研究其对地表水有效性的研究。

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