Evaluation of a PCR detection method for Escherichia coli O157 : H7/H- bovine faecal samples

Al-Ajmi, D., Padmanabha, J., Denman, S. E., Gilbert, R. A., Al Jassim, R. A. M. and McSweeney, C. S. (2006) Evaluation of a PCR detection method for Escherichia coli O157 : H7/H- bovine faecal samples. Letters In Applied Microbiology, 42 4: 386-391. doi:10.1111/j.1472-765X.2005.01850.x


Author Al-Ajmi, D.
Padmanabha, J.
Denman, S. E.
Gilbert, R. A.
Al Jassim, R. A. M.
McSweeney, C. S.
Title Evaluation of a PCR detection method for Escherichia coli O157 : H7/H- bovine faecal samples
Journal name Letters In Applied Microbiology   Check publisher's open access policy
ISSN 0266-8254
1472-765X
Publication date 2006-01-01
Sub-type Article (original research)
DOI 10.1111/j.1472-765X.2005.01850.x
Volume 42
Issue 4
Start page 386
End page 391
Total pages 6
Editor Dr. Jean-Yves Maillard
Place of publication Oxford
Publisher Blackwell Publishing
Collection year 2006
Language eng
Subject C1
300507 Microbiology (excl. Virology)
730216 Food safety
270300 Microbiology
270301 Bacteriology
Formatted abstract
Aims: Combinations of PCR primer sets were evaluated to establish a multiplex PCR method to specifically detect Escherichia coli O157:H7 genes in bovine faecal samples.

Methods and Results: A multiplex PCR method combining three primer sets for the E. coli O157:H7 genes rfbE, uidA and E. coli H7 fliC was developed and tested for sensitivity and specificity with pure cultures of 27 E. coli serotype O157 strains, 88 non-O157 E. coli strains, predominantly bovine in origin and five bacterial strains other than E. coli. The PCR method was very specific in the detection of E. coli O157:H7 and O157:H− strains, and the detection limit in seeded bovine faecal samples was <10 CFU g−1 faeces, following an 18-h enrichment at 37°C, and could be performed using crude DNA extracts as template.

Conclusions: A new multiplex PCR method was developed to detect E. coli O157:H7 and O157:H−, and was shown to be highly specific and sensitive for these strains both in pure culture and in crude DNA extracts prepared from inoculated bovine faecal samples.

Significance and Impact of the Study: This new multiplex PCR method is suitable for the rapid detection of E. coli O157:H7 and O157:H− genes in ruminant faecal samples.
Keyword Bovine Faeces
Escherichia Coli
O157:h7
Pcr Method
Biotechnology & Applied Microbiology
Microbiology
O157
Polymerase-chain-reaction
Multiplex Pcr
Cattle Feces
Genes
Assay
Identification
Amplification
Populations
Prevalence
O157 : H7
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Agriculture and Food Sciences
 
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Created: Wed, 15 Aug 2007, 08:43:42 EST