Detection of pathogenic Escherichia coli on potentially contaminated beef carcasses using cassette PCR and conventional PCR

BMC Microbiology
Dammika P ManageLynn M McMullen

Abstract

Over a one year period, swabs of 820 beef carcasses were tested for the presence of Shiga toxin-producing Escherichia coli by performing Polymerase Chain Reaction (PCR) in a novel technology termed "cassette PCR", in comparison to conventional liquid PCR. Cassette PCR is inexpensive and ready-to-use. The operator need only add the sample and press "go". Cassette PCR can simultaneously test multiple samples for multiple targets. Carcass swab samples were first tested for the presence of STEC genes (O157, eae, stx1 and stx2). Samples were considered to be pathogenic if positive for eae plus stx1 and/or stx2. For samples scored as pathogenic, further testing screened for 6 additional high frequency O-antigens (O26, O45, O103, O111, O121, and O145). Of the 820 samples, 41% were pathogenic and 30% were O157 positive. Of these, 19% of samples were positive for O157 and carried potentially pathogenic E. coli (eae plus stx1 and/or stx2). Of all samples identified as carrying pathogenic E. coli, 18.9, 38.8, 41.4, 0, 36.1, and 4.1% respectively were positive for O26, O45, O103, O111, O121, and O145. To validate cassette PCR testing, conventional PCR using STEC primers was performed on each of the 820 samples. Only 148 of 3280 cassette PC...Continue Reading

References

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Jul 5, 2003·Journal of Clinical Microbiology·Eva Møller Nielsen, Marianne Thorup Andersen
Apr 25, 2007·Critical Reviews in Microbiology·Karl A Bettelheim
Feb 5, 2011·Emerging Infectious Diseases·Ursula KäppeliRoger Stephan
Aug 24, 2013·Lab on a Chip·Dammika P ManageLinda M Pilarski
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Jul 8, 2014·The Journal of Molecular Diagnostics : JMD·Dammika P ManageLinda M Pilarski
May 31, 2015·Methods in Molecular Biology·Dammika P Manage, Linda M Pilarski
Dec 18, 2016·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Mohamed A Karmali
May 11, 2018·PloS One·Darin HuntPatrick M Pilarski

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Methods Mentioned

BETA
PCR
Polymerase Chain Reaction
electrophoresis
PCRs

Software Mentioned

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