Application of mitochondrial DNA analysis for microbial source tracking purposes in shellfish harvesting waters

被引：17

作者：

Baker-Austin, Craig ^{[1
]}

Rangdale, Rachel ^{[1
]}

Lowther, James ^{[1
]}

Lees, David N. ^{[1
]}

机构：

[1] Ctr Environm Fisheries & Aquaculture Sci, Weymouth Lab, Weymouth DT4 8UB, Dorset, England

来源：

WATER SCIENCE AND TECHNOLOGY | 2010年 / 61卷 / 01期

关键词：

faecal contamination; mitochondrial DNA; shellfish; TaqMan; CONTAMINATED SURFACE-WATER; REAL-TIME PCR; FECAL CONTAMINATION; BACTERIA; PORCINE; BOVINE; OVINE;

D O I：

10.2166/wst.2010.767

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

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 (similar to 2 weeks), a similar timeframe of environmental persistence to several bacterial faecal indictors, highlighting its applicability. The procedure described here is specific, rapid (similar to 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.

引用

页码：1 / 7

页数：7

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