Development of a real-time PCR assay with an internal amplification control for the screening of Shiga toxin-producing Escherichia coli in foods.
作者： F. Auvray , C. Lecureuil , F. Dilasser , J. Taché , S. Derzelle
DOI: 10.1111/J.1472-765X.2009.02561.X
关键词:
摘要: 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 Results: A competitive IAC was constructed included stx-specific assay. Coupled to 18-h enrichment automated DNA extraction, could reliably detect presence STEC minced meats inoculated at 10 CFU per 25 g. Its performance evaluated on 415 beef 112 raw milk cheese samples compared with that PCR-ELISA method. Fifty-three 31 cheeses were found stx-positive, giving 98·3% 93·75% concordance, respectively, reference method. Conclusions: highly sensitive method including developed, facilitating monitoring false-negative results due inhibitors. Significance Impact Study: Combined stx-IAC represents rapid foods.
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