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DOI10.1016/j.mimet.2014.07.007
Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods
Haugland, Richard A.1; Siefring, Shawn D.1; Varma, Manju1; Dufour, Alfred P.1; Brenner, Kristen P.1; Wade, Timothy J.2; Sams, Elizabeth2; Cochran, Stacey3; Braun, Steve3; Sivaganensan, Mano4
发表日期2014-10-01
ISSN0167-7012
卷号105页码:59-66
英文摘要

The U.S. EPA has published recommendations for calibrator cell equivalent (CCE) densities of enterococci in recreational waters determined by a qPCR method in its 2012 recreational water quality criteria (RWQC). The CCE quantification unit stems from the calibration model used to estimate enterococci densities in recreational beach waters in the EPA National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study and directly informed the derivation of the RWQC recommendations. Recent studies have demonstrated that CCE estimates from the method can vary when using different cultured Enterococcus cell preparations in calibrator samples. These differences have been attributed to differences in the quantities of targeted gene copies (target sequences) that are recovered per nominal calibrator cell by DNA extraction. Standardization of results from the calibration model will require the estimation of target sequence recoveries from the calibrator and water samples. In addition, comparisons of water sample results with the RWQC values will require a knowledge of target sequence recoveries from the NEEAR study calibrator samples. In this study recoveries of target sequences and the mean target sequence/cell ratio for the NEEAR study calibrator samples were retrospectively estimated with a corroborated standard curve. A modification of the calibration model was then used to estimate enterococci target sequence quantities in water samples from eight midwestem U.S. rivers. CCE estimates were obtained by dividing these target sequence quantities by the mean NEEAR study target sequence/cell ratio. This target sequence-based quantification approach resulted in a high degree of agreement in beach action decisions (determinations of whether bacterial fecal indicator densities are above or below RWQC-recommended values) from CCE results of the qPCR method and from culture dependent enumeration of both enterococci and Eschericia coil in the corresponding water samples. Published by Elsevier B.V.


英文关键词Enterococcus;qPCR;Recreational water quality criteria;EPA methods
语种英语
WOS记录号WOS:000342879500010
来源期刊JOURNAL OF MICROBIOLOGICAL METHODS
来源机构美国环保署
文献类型期刊论文
条目标识符http://gcip.llas.ac.cn/handle/2XKMVOVA/56877
作者单位1.US EPA, Off Res & Dev, Natl Exposure Res Lab, Cincinnati, OH 45268 USA;
2.US EPA, Off Res & Dev, Natl Hlth Effects & Environm Effects Res Lab, Chapel Hill, NC USA;
3.Ohio River Valley Water Sanit Commiss, Cincinnati, OH USA;
4.US EPA, Off Res & Dev, Natl Risk Management Res Lab, Cincinnati, OH 45268 USA
推荐引用方式
GB/T 7714
Haugland, Richard A.,Siefring, Shawn D.,Varma, Manju,et al. Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods[J]. 美国环保署,2014,105:59-66.
APA Haugland, Richard A..,Siefring, Shawn D..,Varma, Manju.,Dufour, Alfred P..,Brenner, Kristen P..,...&Sivaganensan, Mano.(2014).Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods.JOURNAL OF MICROBIOLOGICAL METHODS,105,59-66.
MLA Haugland, Richard A.,et al."Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods".JOURNAL OF MICROBIOLOGICAL METHODS 105(2014):59-66.
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