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Rapid sample preparation method for PCR-based detection of Escherichia coli O157:H7 in ground beef by S. Cui; C.M. Schroeder; D.Y. Zhang; J. Meng is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Aim: To develop an improved, rapid and sensitive sample preparation method for pcr-based detection of escherichia coli o157:h7 in ground beef. ## Methods and results: Fresh ground beef samples were experimentally inoculated with varying concentrations of e. coli o157:h7. pcr inhibitors were removed and bacterial cells were concentrated by filtration and centrifugation, and lysed using enzymatic digestion and successive freeze/thaw cycles. dna was purified and concentrated via phenol/chloroform extraction and the shiga toxin 1 gene (stx1) was amplified using pcr to evaluate the sample preparation method. without prior enrichment of cells in broth media, the detection limit was 103 cfu g-1 beef. when a 6 h enrichment step was incorporated, the detection limit was 1 cfu g-1 beef. the total time required from beginning to end of the procedure was 12 h. ## Conclusions: The sample preparation method developed here enabled substantially improved sensitivity in the pcr-based detection of e. coli o157:h7 in ground beef, as compared to previous reports. ## Significance and impact of the study: Superb sensitivity, coupled with quick turn-around time, relative ease of use and cost-effective

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Author

S. Cui; C.M. Schroeder; D.Y. Zhang; J. Meng

Publisher

Oxford University Press (OUP)

Published

2003

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)