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. 2009 Oct;49(4):529-32.
doi: 10.1111/j.1472-765X.2009.02679.x. Epub 2009 Jul 29.

Rapid identification and differentiation of agricultural faecal contamination sources using multiplex PCR

C Baker-Austin  1 , J Morris, J A Lowther, R Rangdale, D N Lees
Affiliations

Rapid identification and differentiation of agricultural faecal contamination sources using multiplex PCR

C Baker-Austin et al. Lett Appl Microbiol. 2009 Oct.

Abstract

Aims: To develop a quick, easy-to-use, robust and sensitive multiplex PCR assay to detect common sources of agricultural faecal contamination using a combination of bacterial and eukaryote-specific PCR targets.

Method and results: A novel multiplex PCR method was developed that utilizes primers specific for a conserved region of the eukaryote cytochrome-B gene as well as a universal 16S rRNA and the E. coli-specific uidA gene. This multiplex PCR assay was capable of identifying faecal amendments from pig, sheep, cow and goat sources in 24/30 (80%) of amended water samples.

Conclusions: The method was capable of accurately identifying common agricultural sources.

Significance and impact of the study: The procedure described here is simple, rapid (<5 h) and can be used as a first step in microbial source tracking studies, particularly where agricultural faecal contamination is suspected.

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