We present a method for the reliable detection and source characterisation of faecal pollution in water and shellfish matrices, utilising real-time PCR analysis of mitochondrial DNA targets. In this study we designed real-time PCR (TaqMan) probes to target human, bovine, ovine and swine mtDNA. PCR amplification using species-specific TaqMan probes on faecal matter and mixed effluent slurries revealed no cross-reactions between species of interest and other vertebrate faecal matter. Performed as a single blind experiment we were able to correctly identify faecal material in 17/20 effluents (85% correct). mtDNA degrades relatively quickly in faecally-spiked water samples (∼2 weeks), a similar timeframe of environmental persistence to several bacterial faecal indictors, highlighting its applicability. The procedure described here is specific, rapid (<5 hours) and sensitive. These results confirm the suitability of using species-specific mtDNA as an indicator in source tracking studies in surface waters, shellfish harvesting areas and shellfish matrices.
Application of mitochondrial DNA analysis for microbial source tracking purposes in shellfish harvesting waters
Craig Baker-Austin, Rachel Rangdale, James Lowther, David N. Lees; Application of mitochondrial DNA analysis for microbial source tracking purposes in shellfish harvesting waters. Water Sci Technol 1 January 2010; 61 (1): 1–7. doi: https://doi.org/10.2166/wst.2010.767
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