Hill Elizabeth R, Chun Chan Lan, Hamilton Kerry, Ishii Satoshi
Water Resource Science Graduate Program, University of Minnesota, 173 McNeal Hall, 1985 Buford Avenue, St. Paul, Minnesota 55108, United States.
Natural Resources Research Institute, University of Minnesota, 5013 Miller Trunk Highway, Duluth, Minnesota 55811, United States.
ACS ES T Water. 2023 Jul 24;3(8):2647-2658. doi: 10.1021/acsestwater.3c00169. eCollection 2023 Aug 11.
Contamination of water with bacterial, viral, and protozoan pathogens can cause human diseases. Both humans and nonhumans can release these pathogens through their feces. To identify the sources of fecal contamination in the water environment, microbial source tracking (MST) approaches have been developed; however, the relationship between MST markers and pathogens is still not well understood most likely due to the lack of comprehensive datasets of pathogens and MST marker concentrations. In this study, we developed a novel microfluidic quantitative PCR (MFQPCR) platform for the simultaneous quantification of 37 previously validated MST markers, two fecal indicator bacteria (FIB), 22 bacterial, 11 viral, and five protozoan pathogens, and three internal amplification/process controls in many samples. The MFQPCR chip was applied to analyze pathogen removal rates during the wastewater treatment processes. In addition, multiple host-specific MST markers, FIB, and pathogens were successfully quantified in human and avian-impacted surface waters. While the genes for pathogens were relatively infrequently detected, positive correlations were observed between some potential pathogens such as and spp., and human MST markers. The MFQPCR chips developed in this study, therefore, can provide useful information to monitor and improve water quality.
水被细菌、病毒和原生动物病原体污染会导致人类疾病。人类和非人类都可以通过粪便释放这些病原体。为了确定水环境中粪便污染的来源，已经开发了微生物源追踪（MST）方法；然而，MST标记物与病原体之间的关系仍未得到很好的理解，这很可能是由于缺乏病原体和MST标记物浓度的综合数据集。在本研究中，我们开发了一种新型微流控定量PCR（MFQPCR）平台，用于同时定量许多样本中的37种先前验证过的MST标记物、两种粪便指示菌（FIB）、22种细菌、11种病毒和5种原生动物病原体，以及三种内部扩增/过程对照。MFQPCR芯片用于分析废水处理过程中的病原体去除率。此外，在受人类和禽类影响的地表水中成功定量了多种宿主特异性MST标记物、FIB和病原体。虽然病原体基因相对较少被检测到，但在一些潜在病原体（如 和 属）与人类MST标记物之间观察到了正相关。因此，本研究开发的MFQPCR芯片可以为监测和改善水质提供有用信息。
