High-Throughput Microfluidic Quantitative PCR Platform for the Simultaneous Quantification of Pathogens, Fecal...

Area of Science:

• Environmental microbiology
• Water quality monitoring
• Molecular diagnostics

Background:

• Water contamination by bacterial, viral, and protozoan pathogens poses risks to human health.
• Microbial source tracking (MST) methods aim to identify fecal contamination sources, but their link to actual pathogens is unclear due to limited data.
• Existing methods lack the capacity for simultaneous, comprehensive analysis of multiple pathogens and MST markers.

Purpose of the Study:

• To develop a novel microfluidic quantitative PCR (MFQPCR) platform for simultaneous quantification of numerous MST markers, fecal indicator bacteria (FIB), and various pathogens.
• To apply the MFQPCR platform to assess pathogen removal during wastewater treatment.
• To analyze pathogen and MST marker concentrations in human and avian-impacted surface waters.

Main Methods:

• Development of a microfluidic quantitative PCR (MFQPCR) platform.
• Simultaneous quantification of 37 MST markers, 2 FIB, 22 bacterial, 11 viral, and 5 protozoan pathogens.
• Application of the platform to wastewater treatment process samples and surface water samples.

Main Results:

• The MFQPCR platform successfully quantified multiple host-specific MST markers, FIB, and pathogens in various water types.
• Pathogen gene detection was infrequent, but positive correlations were found between specific pathogens (e.g., *Clostridium perfringens*, *Mycobacterium* spp.) and human MST markers.
• The platform demonstrated utility in analyzing pathogen removal rates during wastewater treatment.

Conclusions:

• The developed MFQPCR chip enables comprehensive, simultaneous detection and quantification of waterborne pathogens and MST markers.
• This technology provides valuable data for understanding the relationship between MST markers and pathogens.
• The MFQPCR platform can significantly aid in monitoring and improving water quality by identifying contamination sources.