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Evaluation of rapid qPCR method for quantification of E. coli at non-point source impacted Lake Michigan beaches

Shrestha, Abhilasha, Dorevitch, Samuel
Water research 2019 v.156 pp. 395-403
DNA, Enterococcus, Escherichia coli, United States Environmental Protection Agency, bacteria, beaches, epidemiological studies, indicator species, monitoring, quantitative polymerase chain reaction, water quality, Lake Michigan
Most Great Lakes communities rely on culture-based E. coli methods for monitoring fecal indicator bacteria (FIB) at recreational beaches. These cultivation methods require 18 or more hours to generate results. As a consequence, public notifications about beach action value (BAV) exceedance are based on prior-day water quality. Rapid qPCR monitoring of bacteria in beach water solves the 24-h delay problem, though the USEPA-approved qPCR method targets enterococci bacteria, while Great Lakes communities are familiar with E. coli monitoring. For an E. coli qPCR method to be useful for water quality management, it is important to systematically characterize method performance, and establish BAVs for public notification purposes. In this study, we 1) evaluated a draft USEPA E. coli qPCR method, 2) compared E. coli qPCR measurements with two established FIB (E. coli culture and enterococci qPCR) results, and explored potential strategies to establish E. coli qPCR BAV criteria in the absence of an epidemiological study. Based on analyses of 288 water samples collected from eight of Chicago's Lake Michigan beaches, the E. coli qPCR method demonstrates acceptable performance characteristics. The method is prone to low level DNA contamination, possibly originating from assay reagents derived from E. coli bacteria. Both E. coli and enterococci BAVs were exceeded in approximately 18% of the samples. E. coli qPCR values were correlated with both E. coli culture (r = 0.83; p < 0.0001) and enterococci qPCR (r = 0.67; p < 0.0001) values. The approach recommended by the USEPA in its Technical Support Material (TSM) was used to generate candidate E. coli qPCR BAVs, as was receiver operating characteristic (ROC) analysis. Potential BAV thresholds differed substantially, ranging from 200.9 calibrator cell equivalents (CCE)/100 mL (ROC analysis, enterococci qPCR BAV as the reference) to 1000 CCE/100 mL (TSM analysis, enterococci qPCR BAV as the reference). Because we found that different approaches to establishing potential BAVs generate quite different values, guidance from USEPA about approaches to defining comparable BAVs would be useful.