Publication Abstract

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

Craig Baker-Austin, Joseph Morris, James A. Lowther, Rachel Rangdale, and David N. Lees

Aims

To develop a quick, easy to use, robust and sensitive multiplex PCR assay to detect common sources of agricultural fecal 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 fecal 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 (< 5h) and can be used as a first step in microbial source tracking studies, particularly where agricultural fecal contamination is suspected.

Reference:

Craig Baker-Austin, Joseph Morris, James A. Lowther, Rachel Rangdale, and David N. Lees (2009) Rapid identification and differentiation of agricultural fecal contamination sources using multiplex PCR. Letters in Applied Microbiology 49; 529-532