Comparison of a membrane surface adhesion recovery method with an IMS method for use in a polymerase chain reaction method to detect Escherichia coli O157:H7 in minced beef

Journal of Microbiological Methods
J FitzmauriceM Maher

Abstract

In this study, enrichment procedures and two recovery methods, a membrane surface adhesion technique and an immunomagnetic separation (IMS), were compared for use in conjunction with a multiplex polymerase chain reaction (PCR) method with a view to describing a fast (24 h) and economical test for detection of Escherichia coli O157:H7 in meat samples. The study showed no significant difference between three different enrichment media (BHI, E. coli (E.C.) broth+novobiocin, modified tryptone soya broth (mTSB)+novobiocin) or two incubation temperatures (37 or 41.5 degrees C) for growth of E. coli O157:H7 in minced beef. Minced beef samples inoculated with E. coli O157:H7 at 40 cfu g(-1) were incubated at 37 degrees C for 16 h in E.C. broth+novobiocin reaching numbers of (log(10)7.82-8.70). E. coli O157:H7 were recovered by attachment to polycarbonate membranes immersed in the enriched cultures for 15 min or by immunomagnetic separation. Subsequent treatment of recovered membranes or IMS beads with lysis buffer and phenol/chloroform/isoamyl alcohol was used to extract the DNA from the extracted E. coli O157:H7 cells. The results show when E. coli O157:H7 was present at high levels in the enriched meat sample (log(10)9.6-7.5 cfu ml(-...Continue Reading

References

Mar 24, 1983·The New England Journal of Medicine·L W RileyM L Cohen
Jan 1, 1996·Critical Reviews in Food Science and Nutrition·W E Hill

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Citations

Apr 5, 2011·Meat Science·Rubén GordilloMar Rodríguez
Jan 1, 2008·Meat Science·G DuffyC Cagney
Apr 2, 2015·Critical Reviews in Food Science and Nutrition·Manisha MangalRam K Gupta
Mar 17, 2010·Indian Journal of Dermatology, Venereology and Leprology·Sowmya Kaimal, Devinder Mohan Thappa
Jul 18, 2019·Royal Society Open Science·Baptiste PiqueretPatrizia d'Ettorre
Aug 15, 2017·Brain Pathology·Alexei VerkhratskyVladimir Parpura

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