Application of a rapid qPCR method for enterococci for beach water quality monitoring purposes in Hawaii: Loss of DNA during the extraction protocol due to coral sands

Abstract

Abstract Rapid qPCR methods for enumerating enterococci can provide results in a few hours, thereby enhancing public health protection. Analysis of 140 samples collected from 11 beaches in Hawaii for enterococci using EPA Method 1611 revealed that a majority (70%) of samples yielded unusable data using the recommended protocol due to DNA losses during extraction. The DNA loss was correlated to the presence of suspended coralline sand. Acidification of samples alleviated or removed the interference, enabling successful method application. There were significant correlations across the three methods evaluated (Enterolert®/Method 1600 R2 = 0.85, Enterolert®/ Method 1611 R2 = 0.78 and Method 1600/1611 R2 = 0.82). When samples were not compromised, there was also good agreement among methods for beach management decisions. This study presents a protocol for beach areas with coralline sands, and re-emphasizes the need to use appropriate controls to prevent underestimation of bacterial concentrations at recreational beaches.