Molecular assays for targeting human and bovine enteric viruses in coastal waters and their application for library-independent source tracking
Molecular assays for targeting human and bovine enteric viruses in coastal waters and their application for library-independent source tracking
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DOI：
10.1128/aem.71.4.2070-2078.2005
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发表时间：
2005-04-01
影响因子：
4.4
通讯作者：
Lipp, EK
中科院分区：
文献类型：
--
作者：
Fong, TT;Griffin, DW;Lipp, EK
人口的迅速增长以及沿着水道和沿海地区的城市发展导致水质下降。为了检查上游人为活动对微生物水质的影响，需要采用特定来源的测试方法。在本研究中，开发了针对人肠道病毒（HEV）、牛肠道病毒（BEV）和人腺病毒（HAdV）的分子检测方法，并用于识别格鲁吉亚州阿尔塔马哈河下游粪便污染的主要来源。2002年7月至12月期间，每月从5个受潮汐影响的站点收集2升抓斗样品。通过逆转录和巢式PCR分析样品。PCR结果经斑点杂交证实。在30个地表水样本中，分别有11个和17个样本对HAdV和HEV呈阳性。三分之二的样本对HEV或HAdV检测呈阳性，26%的样本同时出现病毒。在30份地表水样品中，有11份检出BEV。二元逻辑回归分析表明，人类和牛肠道病毒的存在并没有显着相关的粪大肠菌群或总大肠菌群水平。这些病毒的存在是直接相关的溶解氧和流量，但呈负相关的水温，降雨量在30天前采样，叶绿素a浓度。肠道病毒严格的宿主特异性使其成为鉴定水污染源的良好的不依赖于文库的指示物。通过PCR进行病毒病原体检测是一种高度敏感和易于使用的工具，用于在不需要进行公共卫生风险表征时快速评估水质和粪便污染。
Rapid population growth and urban development along waterways and coastal areas have led to decreasing water quality. To examine the effects of upstream anthropogenic activities on microbiological water quality, methods for source-specific testing are required. In this study, molecular assays targeting human enteroviruses (HEV), bovine enteroviruses (BEV), and human adenoviruses (HAdV) were developed and used to identify major sources of fecal contamination in the lower Altamaha River, Georgia. Two-liter grab samples were collected monthly from five tidally influenced stations between July and December 2002. Samples were analyzed by reverse transcription- and nested-PCR. PCR results were confirmed by dot blot hybridization. Eleven and 17 of the 30 surface water samples tested positive for HAdV and HEV, respectively. Two-thirds of the samples tested positive for either HEV or HAdV, and the viruses occurred simultaneously in 26% of samples. BEV were detected in 11 of 30 surface water samples. Binary logistic regression analysis showed that the presence of both human and bovine enteric viruses was not significantly related to either fecal coliform or total coliform levels. The presence of these viruses was directly related to dissolved oxygen and streamflow but inversely related to water temperature, rainfall in the 30 days preceding sampling, and chlorophyll-a concentrations. The stringent host specificity of enteric viruses makes them good library-independent indicators for identification of water pollution sources. Viral pathogen detection by PCR is a highly sensitive and easy-to-use tool for rapid assessment of water quality and fecal contamination when public health risk characterization is not necessary.