A real-time polymerase chain reaction assay for quantitative detection of the human-specific enterococci surface protein marker in sewage and environmental waters

Environmental Microbiology
W AhmedD Powell

Abstract

A real-time polymerase chain reaction (PCR) assay using SYBR Green I dye was developed to quantify the Enterococcus faecium enterococci surface protein (esp) marker in sewage (n = 16) and environmental waters (n = 16). The concentration of culturable enterococci in raw sewage samples ranged between 1.3 x 10(5) and 5.6 x 10(5) colony-forming units (cfu) per 100 ml. The real-time PCR detected 9.8 x 10(3)-3.8 x 10(4) gene copies of the esp marker per 100 ml of sewage. However, the concentration of culturable enterococci and the esp marker in secondary effluent was two orders of magnitude lower than raw sewage. Surface water samples were collected from a non-sewered catchment after storm events and the real-time PCR was applied to quantify the esp marker. Of the 16 samples tested, 6 (38%) were PCR-positive and the concentration of the esp marker ranged between 1.1 x 10(2) and 5.3 x 10(2) gene copies per 100 ml of water samples. The newly developed real-time PCR method was successfully used to quantify the esp marker in samples collected from sewage and environmental waters. The presence of the esp marker in water samples immediately after storm events not only indicated human faecal pollution but also provided evidence of the degre...Continue Reading

References

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Citations

Jan 7, 2011·Applied Microbiology and Biotechnology·Peter Roslev, Annette S Bukh
Dec 4, 2012·Microbiology and Molecular Biology Reviews : MMBR·Muruleedhara N ByappanahalliValerie J Harwood
May 22, 2009·Journal of Applied Microbiology·A KorajkicV J Harwood
Oct 30, 2010·Journal of Applied Microbiology·A KorajkicV J Harwood
Sep 21, 2011·Environmental Monitoring and Assessment·Kelvin Wong, Irene Xagoraraki
Sep 20, 2008·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Gauri R VaradhacharyJohn D Hainsworth
Jun 28, 2019·Applied Microbiology and Biotechnology·Carmen Astudillo-GarcíaGavin Lear
Jun 29, 2021·Frontiers in Microbiology·Silvia MonteiroRicardo Santos

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