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J Microbiol Methods. 2020 Dec;179:106086. doi: 10.1016/j.mimet.2020.106086. Epub 2020 Oct 13.
2
Simplified Analysis of Measurement Data from A Rapid qPCR Method (EPA Draft Method C) Using A Standardized Excel Workbook.使用标准化Excel工作簿对快速定量聚合酶链反应方法(美国环境保护局草案方法C)的测量数据进行简化分析
Water (Basel). 2020 Mar 11;12(3):1-775. doi: 10.3390/w12030775.
3
Evaluation of multiple laboratory performance and variability in analysis of recreational freshwaters by a rapid Escherichia coli qPCR method (Draft Method C).应用快速大肠杆菌 qPCR 方法(草案方法 C)评估娱乐性淡水中多个实验室的检测性能和分析变异性。
Water Res. 2019 Jun 1;156:465-474. doi: 10.1016/j.watres.2019.03.014. Epub 2019 Mar 16.
4
Standardized data quality acceptance criteria for a rapid Escherichia coli qPCR method (Draft Method C) for water quality monitoring at recreational beaches.用于海滩水质监测的快速大肠杆菌 qPCR 方法(草案方法 C)的数据质量标准化验收标准。
Water Res. 2019 Jun 1;156:456-464. doi: 10.1016/j.watres.2019.03.011. Epub 2019 Mar 15.
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Advancements in mitigating interference in quantitative polymerase chain reaction (qPCR) for microbial water quality monitoring.在减轻定量聚合酶链反应(qPCR)中微生物水质监测干扰方面的进展。
Sci Total Environ. 2019 Jun 25;671:732-740. doi: 10.1016/j.scitotenv.2019.03.242. Epub 2019 Mar 16.
6
Evaluation of rapid qPCR method for quantification of E. coli at non-point source impacted Lake Michigan beaches.评价快速 qPCR 方法在非点源污染密西根湖海滩中定量检测大肠杆菌的效果。
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J Environ Qual. 2018 Sep;47(5):1086-1093. doi: 10.2134/jeq2017.11.0462.
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Quantification of plasmid DNA standards for U.S. EPA fecal indicator bacteria qPCR methods by droplet digital PCR analysis.通过滴液数字PCR分析对美国环保署粪便指示菌qPCR方法的质粒DNA标准品进行定量。
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10
Monitoring urban beaches with qPCR vs. culture measures of fecal indicator bacteria: Implications for public notification.采用定量聚合酶链反应(qPCR)与粪便指示菌培养法监测城市海滩:对公共通报的影响
Environ Health. 2017 May 12;16(1):45. doi: 10.1186/s12940-017-0256-y.

在密歇根州实施快速、多点海滩检测的过程中,对基于培养法和 qPCR 法(EPA 草案方法 C)检测的大肠杆菌水平进行大规模比较。

Large-scale comparison of E. coli levels determined by culture and a qPCR method (EPA Draft Method C) in Michigan towards the implementation of rapid, multi-site beach testing.

机构信息

U.S. Environmental Protection Agency, Office of Research and Development, Center for Environmental Measurements and Modeling, Cincinnati, OH, United States of America.

U.S. Environmental Protection Agency, Office of Research and Development, Center for Environmental Measurements and Modeling, Cincinnati, OH, United States of America.

出版信息

J Microbiol Methods. 2021 May;184:106186. doi: 10.1016/j.mimet.2021.106186. Epub 2021 Mar 22.

DOI:10.1016/j.mimet.2021.106186
PMID:33766609
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8650687/
Abstract

Fecal pollution remains a challenge for water quality managers at Great Lakes and inland recreational beaches. The fecal indicator of choice at these beaches is typically Escherichia coli (E. coli), determined by culture-based methods that require over 18 h to obtain results. Researchers at the United States Environmental Protection Agency (EPA) have developed a rapid E. coli qPCR methodology (EPA Draft Method C) that can provide same-day results for improving public health protection with demonstrated sensitivity, specificity, and data acceptance criteria. However, limited information is currently available to compare the occurrence of E. coli determined by cultivation and by EPA Draft Method C (Method C). This study provides a large-scale data collection effort to compare the occurrence of E. coli determined by these alternative methods at more than 100 Michigan recreational beach and other sites using the complete set of quantitative data pairings and selected subsets of the data and sites meeting various eligibility requirements. Simple linear regression analyses of composite (pooled) data indicated a correlation between results of the E. coli monitoring approaches for each of the multi-site datasets as evidenced by Pearson R-squared values ranging from 0.452 to 0.641. Theoretical Method C threshold values, expressed as mean log10 target gene copies per reaction, that corresponded to an established E. coli culture method water quality standard of 300 MPN or CFU /100 mL varied only from 1.817 to 1.908 for the different datasets using this model. Different modeling and derivation approaches that incorporated within and between-site variability in the estimates also gave Method C threshold values in this range but only when relatively well-correlated datasets were used to minimize the error. A hypothetical exercise to evaluate the frequency of water impairments based on theoretical qPCR thresholds corresponding to the E. coli water quality standard for culture methods suggested that the methods may provide the same beach notification outcomes over 90% of the time with Method C results differing from culture method results that indicated acceptable and unacceptable water quality at overall rates of 1.9% and 6.6%, respectively. Results from this study provide useful information about the relationships between E. coli determined by culture and qPCR methods across many diverse freshwater sites and should facilitate efforts to implement qPCR-based E. coli detection for rapid recreational water quality monitoring on a large scale in the State of Michigan.

摘要

粪便污染仍然是大湖和内陆休闲海滩水质管理人员面临的挑战。这些海滩上选择的粪便指标通常是大肠杆菌(E. coli),通过基于培养的方法确定,这些方法需要超过 18 小时才能获得结果。美国环境保护署(EPA)的研究人员开发了一种快速大肠杆菌 qPCR 方法(EPA 草案方法 C),该方法可以提供当日结果,提高公共卫生保护水平,具有证明的敏感性、特异性和数据接受标准。然而,目前可用的信息有限,无法比较培养法和 EPA 草案方法 C(方法 C)确定的大肠杆菌的发生情况。本研究提供了一项大规模的数据收集工作,使用完整的定量数据配对和符合各种资格要求的数据和地点的选定子集,比较了密歇根州 100 多个休闲海滩和其他地点使用这些替代方法确定的大肠杆菌的发生情况。对每个多地点数据集的大肠杆菌监测方法的结果进行简单线性回归分析,表明了复合(合并)数据中 Pearson R-squared 值从 0.452 到 0.641 的相关性,表明这些结果存在相关性。理论上的方法 C 阈值值,以每个反应的目标基因拷贝的平均对数 10 表示,与既定的大肠杆菌培养方法水质标准 300 MPN 或 CFU /100 相比,仅在使用该模型的不同数据集之间变化为 1.817 到 1.908。该模型还采用了包含和站点之间变异性的不同建模和推导方法,给出了该范围内的方法 C 阈值值,但只有当使用相关性较好的数据集来最小化误差时,才会给出方法 C 阈值值。根据与培养方法大肠杆菌水质标准相对应的理论 qPCR 阈值评估水质受损频率的假设性练习表明,在 90%以上的时间内,方法 C 的结果可能会提供相同的海滩通知结果,而方法 C 的结果与表明可接受和不可接受水质的培养方法结果的差异率分别为 1.9%和 6.6%。本研究的结果提供了有关培养法和 qPCR 法在许多不同淡水场所确定的大肠杆菌之间关系的有用信息,应有助于在密歇根州大规模实施基于 qPCR 的大肠杆菌检测,用于快速休闲水质监测。