Development of a qPCR assay for the detection of naturalized wastewater E. coli strains

We recently demonstrated the presence of naturalized populations of Escherichia coli in municipal sewage. We wanted to develop a quantitative polymerase chain reaction (qPCR) assay targeting the uspC-IS30-flhDC marker of naturalized wastewater E. coli and assess the prevalence of these naturalized s...

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Published in:Journal of water and health 2022-04, Vol.20 (4), p.727-736
Main Authors: Zhi, Shuai, Banting, Graham, Neumann, Norman F
Format: Article
Language:English
Subjects:
Animals
Assaying
Bacteroides
Biomarkers
Cloning
Contamination
Detection
DNA
E coli
Effluent treatment
Effluents
Escherichia coli - genetics
Fecal coliforms
Feces
Genomes
Heat resistance
Human wastes
Municipal wastes
Municipal wastewater
naturalized
Nucleotide sequence
PCR
Plasmids
Polymerase chain reaction
Public health
qpcr
Sewage
Sewage - analysis
Sewage disposal
Specificity
Stormwater
Surface water
Waste Water - analysis
wastewater contamination
Wastewater pollution
Wastewater treatment
Water pollution
Water Purification
Water quality
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description We recently demonstrated the presence of naturalized populations of Escherichia coli in municipal sewage. We wanted to develop a quantitative polymerase chain reaction (qPCR) assay targeting the uspC-IS30-flhDC marker of naturalized wastewater E. coli and assess the prevalence of these naturalized strains in wastewater. The limit of detection for the qPCR assay was 3.0 × 10 ng of plasmid DNA template with 100% specificity. This strain was detected throughout the wastewater treatment process, including treated effluents. We evaluated the potential of this marker for detecting municipal sewage/wastewater contamination in water by comparing it to other human and animal markers of fecal pollution. Strong correlations were observed between the uspC-IS30-flhDC marker and the human fecal markers Bacteroides HF183 and HumM2, but not animal fecal markers, in surface and stormwater samples. The uspC-IS30-flhDC marker appears to be a potential E. coli-based marker for human wastewater contamination.
doi_str_mv 10.2166/wh.2022.014
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subjects Animals
Assaying
Bacteroides
Biomarkers
Cloning
Contamination
Detection
DNA
E coli
Effluent treatment
Effluents
Escherichia coli - genetics
Fecal coliforms
Feces
Genomes
Heat resistance
Human wastes
Municipal wastes
Municipal wastewater
naturalized
Nucleotide sequence
PCR
Plasmids
Polymerase chain reaction
Public health
qpcr
Sewage
Sewage - analysis
Sewage disposal
Specificity
Stormwater
Surface water
Waste Water - analysis
wastewater contamination
Wastewater pollution
Wastewater treatment
Water pollution
Water Purification
Water quality
title Development of a qPCR assay for the detection of naturalized wastewater E. coli strains
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