TY - JOUR
T1 - High-Throughput Microfluidic Quantitative PCR Platform for the Simultaneous Quantification of Pathogens, Fecal Indicator Bacteria, and Microbial Source Tracking Markers
AU - Hill, Elizabeth R.
AU - Chun, Chan Lan
AU - Hamilton, Kerry
AU - Ishii, Satoshi
N1 - Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.
PY - 2023/8/11
Y1 - 2023/8/11
N2 - 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 Clostridium perfringens and Mycobacterium spp., and human MST markers. The MFQPCR chips developed in this study, therefore, can provide useful information to monitor and improve water quality.
AB - 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 Clostridium perfringens and Mycobacterium spp., and human MST markers. The MFQPCR chips developed in this study, therefore, can provide useful information to monitor and improve water quality.
KW - fecal pollution
KW - microbial source tracking
KW - pathogens
KW - quantitative PCR
KW - water quality
UR - http://www.scopus.com/inward/record.url?scp=85167895043&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85167895043&partnerID=8YFLogxK
U2 - 10.1021/acsestwater.3c00169
DO - 10.1021/acsestwater.3c00169
M3 - Article
C2 - 37593240
AN - SCOPUS:85167895043
SN - 2690-0637
VL - 3
SP - 2647
EP - 2658
JO - ACS ES and T Water
JF - ACS ES and T Water
IS - 8
ER -