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


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.


CONCLUSIONS
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.
Baker-Austin
C
C
Centre for Environment, Fisheries and Aquaculture Science, Weymouth Laboratory, Weymouth, Dorset, UK. craig.baker-austin@cefas.co.uk
Morris
J
J
Lowther
J A
JA
Rangdale
R
R
Lees
D N
DN
D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques / Q000379:methods*; D045303:Cytochromes b / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D045303:Cytochromes b