Abstract:
AIMS:To develop and evaluate a real-time PCR assay incorporating an internal amplification control (IAC) suitable for the screening of Shiga toxin (Stx)-producing Escherichia coli (STEC) in foods. METHODS AND RESULTS:A competitive IAC was constructed and included in an stx-specific real-time PCR assay. Coupled to 18-h enrichment and automated DNA extraction, the assay could reliably detect the presence of STEC in minced meats inoculated at 10 CFU per 25 g. Its performance was evaluated on 415 minced beef and 112 raw milk cheese samples and compared with that of a PCR-ELISA method. Fifty-three minced meats and 31 cheeses were found stx-positive, giving 98.3% and 93.75% concordance, respectively, with the PCR-ELISA reference method. CONCLUSIONS:A highly sensitive stx-specific real-time PCR method including an IAC was developed, facilitating monitoring of false-negative results due to PCR inhibitors. SIGNIFICANCE AND IMPACT OF THE STUDY:Combined with automated DNA extraction, the stx-IAC real-time PCR assay represents a suitable method for rapid screening of STEC in foods.
journal_name
Lett Appl Microbioljournal_title
Letters in applied microbiologyauthors
Auvray F,Lecureuil C,Dilasser F,Taché J,Derzelle Sdoi
10.1111/j.1472-765X.2009.02561.xsubject
Has Abstractpub_date
2009-05-01 00:00:00pages
554-9issue
5eissn
0266-8254issn
1472-765Xpii
LAM2561journal_volume
48pub_type
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