A real-time polymerase chain reaction assay for the quantitative detection of the human-specific enterococci surface protein marker in sewage and environmental waters
Ahmed, Warish, Stewart, Joel, Gardner, Ted, & Powell, Daniel (2008) A real-time polymerase chain reaction assay for the quantitative detection of the human-specific enterococci surface protein marker in sewage and environmental waters. Environmental Microbiology, 10(12), pp. 3255-3264.
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 degree of human faecal pollution. To our knowledge, this is the first study that reports the use of a real-time PCR assay to quantify the esp marker in sewage and surface waters. Such study would provide valuable information for managers for the improved management of water quality.
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|Item Type:||Journal Article|
|Keywords:||Microbial source tracking, Faecal indicators, water pollution|
|Divisions:||Past > QUT Faculties & Divisions > Faculty of Built Environment and Engineering|
|Copyright Owner:||Copyright 2008 Blackwell Publishing|
|Copyright Statement:||Self-archiving of the author-version in institutional repositories is not currently supported by this publisher. For more information, please refer to the journal's website (see hypertext link) or contact the author.|
|Deposited On:||24 Feb 2009 21:46|
|Last Modified:||10 Aug 2011 17:38|
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