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Immunomagnetic separation as a sensitive method for isolating Escherichia coli O157 from food samples

  • D. J. Wright (a1), P. A. Chapman (a1) and C. A. Siddons (a1)
  • DOI: http://dx.doi.org/10.1017/S0950268800051438
  • Published online: 01 May 2009
Abstract
SUMMARY

Minced beef samples inoculated with Escherichia coli O157 were cultured in buffered peptone water supplemented with vancomycin, cefsulodin and cefixime (BPW-VCC) and subcultured to cefixime tellurite sorbitol MacConkey (CT-SMAC) agar both directly and after immunomagnetic separation (IMS) of the organism with magnetic beads coated with an antibody against E. coli O157 (Dynabeads anti-E. coli 0157, Dynal, Oslo). E. coli O157 was recovered from initial inocula of 200 organisms/g by direct subculture and 2 organisms/g by IMS. Twelve strains of E. coli 0157 of different combinations of phage type, H antigen and toxin genotype were all recovered from initial inocula of two organisms/g by IMS. Nonspecific binding of other organisms to the magnetic beads could be reduced by washing of the beads in PBS with Tween-20 0·002–0.005% E. coli O157 was not bound by magnetic coated with an unrelated antibody.

During investigation of a dairy herd that was possibly linked to a small outbreak of infection with E. coli O157, the organism was isolated from 2 of 279 forestream milk samples from individual cattle; both isolates were made only by the IMS technique. IMS is rapid, technically simple, and a specific method for isolation of E. coli O157 and will be useful in epidemiological studies.

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7.PA Chapman , CA Siddons , PM Zadik , L Jewes . An improved selective medium for the isolation of Escherichia coli O157. J Med Microbiol 1991; 35: 107–10.

8.PM Zadik , PA Chapman , CA Siddons . Use of tellurite for the selection of verocytotoxigenic Escherichia coli O157. J Med Microbiol 1993; 39: 155–8.

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Epidemiology & Infection
  • ISSN: 0950-2688
  • EISSN: 1469-4409
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