Southern California Coastal Water Research Project Authority, Costa Mesa, CA 92626, USA.
U.S. Environmental Protection Agency, Office of Research and Development, Cincinnati, OH 45268, USA.
Water Res. 2018 Jan 1;128:148-156. doi: 10.1016/j.watres.2017.10.071. Epub 2017 Oct 31.
Human fecal pollution of recreational waters remains a public health concern worldwide. As a result, there is a growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for water quality research and management. However, there are currently no standardized approaches for field implementation and interpretation of qPCR data. In this study, a standardized HF183/BacR287 qPCR method was combined with a water sampling strategy and a novel Bayesian weighted average approach to establish a human fecal contamination score (HFS) that can be used to prioritize sampling sites for remediation based on measured human waste levels. The HFS was then used to investigate 975 study design scenarios utilizing different combinations of sites with varying sampling intensities (daily to once per week) and number of qPCR replicates per sample (2-14 replicates). Findings demonstrate that site prioritization with HFS is feasible and that both sampling intensity and number of qPCR replicates influence reliability of HFS estimates. The novel data analysis strategy presented here provides a prescribed approach for the implementation and interpretation of human-associated HF183/BacR287 qPCR data with the goal of site prioritization based on human fecal pollution levels. In addition, information is provided for future users to customize study designs for optimal HFS performance.
人类粪便污染休闲水域仍然是全球公共卫生关注的问题。因此,人们越来越关注将人类相关粪便源识别定量实时 PCR(qPCR)技术应用于水质研究和管理。然而,目前还没有标准化的方法来进行现场实施和解释 qPCR 数据。在这项研究中,标准化的 HF183/BacR287 qPCR 方法与一种水样采集策略和一种新的贝叶斯加权平均方法相结合,建立了一个人类粪便污染评分(HFS),可用于根据测量的人类废物水平优先选择修复采样点。然后,使用 HFS 研究了 975 种不同的研究设计方案,这些方案结合了不同采样强度(每天至每周一次)和每个样品的 qPCR 重复次数(2-14 次)的不同采样点组合。研究结果表明,使用 HFS 进行站点优先级排序是可行的,并且采样强度和 qPCR 重复次数都会影响 HFS 估计的可靠性。这里提出的新数据分析策略为实施和解释基于人类粪便污染水平的人类相关 HF183/BacR287 qPCR 数据提供了一种规定的方法。此外,还为未来的用户提供了定制研究设计以获得最佳 HFS 性能的信息。