TY - JOUR
T1 - Rapid identification and differentiation of agricultural faecal contamination sources using multiplex PCR
AU - Baker-Austin, C.
AU - Morris, J.
AU - Lowther, J. A.
AU - Rangdale, R.
AU - Lees, D. N.
PY - 2009/10
Y1 - 2009/10
N2 - 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.
AB - 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.
KW - Faecal contamination
KW - Microbial source-tracking
KW - Mitochondrial DNA
KW - Multiplex PCR
UR - http://www.scopus.com/inward/record.url?scp=70349114573&partnerID=8YFLogxK
U2 - 10.1111/j.1472-765X.2009.02679.x
DO - 10.1111/j.1472-765X.2009.02679.x
M3 - Article
C2 - 19708886
AN - SCOPUS:70349114573
SN - 0266-8254
VL - 49
SP - 529
EP - 532
JO - Letters in Applied Microbiology
JF - Letters in Applied Microbiology
IS - 4
ER -