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

MARINE POLLUTION BULLETIN

卷:149

Article

Seruge, Jaline¹  Wong, Mayee¹  Noble, Rachel T.²  Blackwood, A. Denene²  Moravcik, Philip S.¹  Kirs, Marek¹ 

[1] Univ Hawaii, Water Resources Res Ctr, 2540 Dole St,Holmes Hall 283, Honolulu, HI 96822 USA

[2] Univ North Carolina Chapel Hill, Inst Marine Sci, 3431 Arendell St, Morehead City, NC 28557 USA

关键词: Enterococci;    Coastal water quality;    EPA method 1611;    Interference;    Hawaii;   

DOI  :  10.1016/j.marpolbul.2019.110631

来源: Elsevier

【 摘 要 】

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 (R)/Method 1600 R-2 = 0.85, Enterolert (R)/Method 1611 R-2 = 0.78 and Method 1600/1611 R-2 = 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.

【 授权许可】

Free