Development of a qPCR assay for the detection of naturalized wastewater E. coli strains. (12th April 2022)

• Record Type: Journal Article
• Authors: Zhi, Shuai; Banting, Graham; Neumann, Norman F.
• Is Part Of: Journal of water and health. Volume 20:Number 4(2022)
• Page Start: 727
• Page End: 736
• Publication Date: 2022-04-12
• Subjects: E. coli -- naturalized -- qPCR -- wastewater contamination; Water quality management -- Periodicals; Water -- Pollution -- Environmental aspects -- Periodicals; Environmental health -- Periodicals; Water quality -- Health aspects -- Periodicals; Water -- Health aspects -- Periodicals; Water -- Pollution -- Health aspects -- Periodicals; Public Health; Water Pollution -- prevention & control; Quality Control; Water Microbiology; Water Supply -- standards; Health & Medicine (General); Hydrology; Environmental health; Water -- Health aspects; Water -- Pollution -- Environmental aspects; Water -- Pollution -- Health aspects; Water quality -- Health aspects; Water quality management; Water; Gezondheid; Periodical; Periodicals; 363.61
• DOI: 10.2166/wh.2022.014
• Languages: English
• ISSNs: 1477-8920

Abstract

Abstract: 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 −8 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. HIGHLIGHTS: A quantitative polymerase chain reaction assay of naturalized wastewater Escherichia coli was successfully developed with a low limit of detection and 100% specificity. 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. Graphical Abstract