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Molecular Method for Detection of Total Coliforms in Drinking Water Samples

机译:饮用水样品中总大肠菌群的分子检测方法

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摘要

This work demonstrates the ability of a bacterial concentration and recovery procedure combined with three different PCR assays targeting the lacZ, wecG, and 16S rRNA genes, respectively, to detect the presence of total coliforms in 100-ml samples of potable water (presence/absence test). PCR assays were first compared to the culture-based Colilert and MI agar methods to determine their ability to detect 147 coliform strains representing 76 species of Enterobacteriaceae encountered in fecal and environmental settings. Results showed that 86 (58.5%) and 109 (74.1%) strains yielded a positive signal with Colilert and MI agar methods, respectively, whereas the lacZ, wecG, and 16S rRNA PCR assays detected 133 (90.5%), 111 (75.5%), and 146 (99.3%) of the 147 total coliform strains tested. These assays were then assessed by testing 122 well water samples collected in the Québec City region of Canada. Results showed that 97 (79.5%) of the samples tested by culture-based methods and 95 (77.9%), 82 (67.2%), and 98 (80.3%) of samples tested using PCR-based methods contained total coliforms, respectively. Consequently, despite the high genetic variability of the total coliform group, this study demonstrated that it is possible to use molecular assays to detect total coliforms in potable water: the 16S rRNA molecular assay was shown to be as efficient as recommended culture-based methods. This assay might be used in combination with an Escherichia coli molecular assay to assess drinking water quality.
机译:这项工作证明了细菌浓缩和回收程序与分别针对lacZ,wecG和16S rRNA基因的三种不同PCR测定法相结合的能力,能够检测100 ml饮用水样品中总大肠菌的存在(存在/不存在)测试)。首先将PCR检测方法与基于培养物的Collilert和MI琼脂方法进行比较,以确定它们检测在粪便和环境中遇到的代表76种肠杆菌科细菌的147种大肠杆菌的能力。结果显示,分别用Colilert和MI琼脂方法分别对86(58.5%)和109(74.1%)菌株产生了阳性信号,而lacZ,wecG和16S rRNA PCR检测法分别检测到133(90.5%),111(75.5%) )和147种测试的大肠菌中的146种(99.3%)。然后通过测试在加拿大魁北克市地区收集的122口井水样品来评估这些测定。结果显示,通过基于培养的方法测试的样品中有97(79.5%),使用基于PCR的方法测试的样品中分别有95(77.9%),82(67.2%)和98(80.3%)包含大肠菌群。因此,尽管总大肠菌群的遗传变异性很高,但这项研究表明,可以使用分子测定法来检测饮用水中的总大肠菌群:16S rRNA分子测定法与推荐的基于培养的方法一样有效。该测定法可与大肠杆菌分子测定法结合使用以评估饮用水质量。

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