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

Published online by Cambridge University Press:  15 May 2009

D. J. Wright ,
P. A. Chapman  and
C. A. Siddons
D. J. Wright
Affiliation:
Public Health Laboratory, Herries Road, Sheffield S5 7BQ
P. A. Chapman*
Affiliation:
Public Health Laboratory, Herries Road, Sheffield S5 7BQ
C. A. Siddons
Affiliation:
Public Health Laboratory, Herries Road, Sheffield S5 7BQ
*
* Author for correspondence.
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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.

Type
Research Article
Information
Epidemiology & Infection , Volume 113 , Issue 1 , August 1994 , pp. 31 - 39
Copyright
Copyright © Cambridge University Press 1994

References

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