Hostname: page-component-586b7cd67f-tf8b9 Total loading time: 0 Render date: 2024-11-22T05:04:57.688Z Has data issue: false hasContentIssue false
  • English
  • Français

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.
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

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

REFERENCES

1.Riley, LW, Remis, RS, Helgerson, SD et al. , Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med 1983; 308: 681–5.CrossRefGoogle ScholarPubMed
2.Martin, ML, Shipman, LD, Wells, JG et al. , Isolation of Escherichia coli 0157: H7 from dairy cattle associated with two cases of haemolytic-uraemic syndrome. Lancet 1986; 2: 1043.CrossRefGoogle Scholar
3.Chapman, PA, Wright, DJ, Norman, P. Verotoxin-producing Escherichia coli infections in Sheffield: cattle as a possible source. Epidemiol Infect 1989; 102: 439–45.CrossRefGoogle ScholarPubMed
4.Chapman, PA, Siddons, CA, Wright, DJ, Norman, P, Fox, J, Crick, E. Cattle as a possible source of verocytotoxin-producing Escherichia coli O157 infections in man. Epidemiol Infect 1993; 111: 439–47.CrossRefGoogle ScholarPubMed
5.Chapman, PA, Wright, DJ, Higgins, R. Untreated milk as a source of verotoxigenic E. coli 0157. Vet Rec 1993; 133: 171–2CrossRefGoogle Scholar
6.March, SB, Ratnam, S. Sorbitol-MacConkey medium for the detection of Escherichia coli 0157:H7 associated with hemorrhagic colitis. J Clin Microbiol 1986; 23: 869–72.CrossRefGoogle Scholar
7.Chapman, PA, Siddons, CA, Zadik, PM, Jewes, L. An improved selective medium for the isolation of Escherichia coli O157. J Med Microbiol 1991; 35: 107–10.CrossRefGoogle ScholarPubMed
8.Zadik, PM, Chapman, PA, Siddons, CA. Use of tellurite for the selection of verocytotoxigenic Escherichia coli O157. J Med Microbiol 1993; 39: 155–8.CrossRefGoogle ScholarPubMed
9.Sambrook, J, Fritsch, EF, Maniatis, T. Molecular cloning. 2nd ed.Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1989: E1–E2.Google Scholar
10.Chapman, PA. Evaluation of a commercial latex slide test for the identification of Escherichia coli 0157. J Clin Pathol 1989; 42: 1109–10.CrossRefGoogle Scholar
11.Borczyk, AA, Karmali, MA, Lior, H, Duncan, LM. Bovine reservoir for verotoxin-producing Escherichia coli 0157:H7. Lancet 1987; 1: 98.CrossRefGoogle Scholar
12.Ostroff, SM. Griffin, PM. Tauxe, RV et al. , A statewide outbreak of Escherichia coli O157 infections in Washington state. Am J Epidemiol 1990; 132: 239–47.CrossRefGoogle ScholarPubMed
13.Wells, JG, Shipman, LD. Greene, KD et al. , Isolation of Escherichia coli serotype O157:H7 and other shiga-like toxin-producing E. coli from dairy cattle. J Clin Microbiol 1991; 29: 985–9.CrossRefGoogle ScholarPubMed
14.Synge, BA, Hopkins, GF. Verotoxigenic Escherichia coli O157 in Scottish calves. Vet Rec 1992: 130: 583.CrossRefGoogle Scholar
15.Raghubeer, EV, Matches, JR. Temperature range for growth of Escherichia coli O157:H7 and selected coliforms in E. coli medium. J Clin Microbiol 1990; 28: 803–5.CrossRefGoogle ScholarPubMed
16.Doyle, MP, Schoeni, JL. Survival and growth characteristics of Escherichia coli associated with hemorrhagic colitis. Appl Environ Microbiol 1984; 48: 855–6.CrossRefGoogle ScholarPubMed
17.Fratamico, PM, Schultz, FJ, Buchanan, RL. Rapid isolation of Escherichia coli O157:H7 from enrichment cultures of foods using an immunomagnetic separation method. Food Microbiol 1992; 9: 105–13.CrossRefGoogle Scholar
18.Okrend, AJG, Bonnie, ER, Lattuada, CP. Isolation of Escherichia coli O157:H7 using O157 specific antibody coated magnetic beads. J Food Protect 1992; 55: 214–17.CrossRefGoogle Scholar
19.Padhye, NV, Doyle, MP. Production and characterization of a monoclonal antibody specific for enterohemorrhagic Escherichia coli of serotypes O157:H7 and 026: H11. J Clin Microbiol 1991; 29: 99–103.CrossRefGoogle Scholar
20.Sernowski, LP, Ingham, SC. Frequency of false presumptive positive results obtained using a commercial ELISA kit to screen retail ground beef for Escherichia coli 0157: H7. J Food Protect 1992; 55: 846.Google Scholar
21.Szabo, RAE, Todd, E, MacKenzie, J, Parrington, L, Armstrong, A. Increased sensitivity of the rapid hydrophobic grid membrane filter enzyme labelled antibody procedure for Escherichia coli O157 detection in foods and bovine feces. Appl Environ Microbiol 1990; 56: 3546–9.CrossRefGoogle ScholarPubMed
22.Doyle, MP, Schoeni, JL. Isolation of Escherichia coli O157:H7 from retail fresh meats and poultry. Appl Environ Microbiol 1987; 83: 2394–6.CrossRefGoogle Scholar
23.Gannon, VPJ, King, RK, Kim, JY, Golsteyn, Thomas EJ. Rapid and sensitive method for detection of shiga-like toxin-producing Escherichia coli ground beef using the polymerase chain reaction. Appl Environ Microbiol 1992; 58: 3809–15.CrossRefGoogle ScholarPubMed
24.Read, SC, Gyles, CL, Clarke, RC, Lior, H, McEwen, S. Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork and chicken in southwestern Ontario. Epidemiol Infect 1990: 105: 11–20.CrossRefGoogle ScholarPubMed
25.Clarke, RC, McEwen, SA, Gannon, VP, Lior, H, Gyles, CL. Isolation of verocytotoxin-producing Escherichia coli from milk filters in south-western Ontario. Epidemiol Infect 1989; 102: 253–60.CrossRefGoogle ScholarPubMed
26.Roberts, D, Thirlwell, J, Smith, H, Willshaw, G. The microbiological examination of raw beef products with particular reference to Vero cytotoxin producing Escherichia coli. Proceedings of the 3rd World Congress on Foodborne Infections and Intoxications, 06 1992, Berlin, Volume 1, 506–508. Institute of Veterinary Medicine, Berlin.Google Scholar