1887

Abstract

Summary

Shiga toxin and Shiga-like toxins (SLTs, syn. Verotoxins) are currently detected by tissue culture assays that are expensive, time-consuming and require specialised facilities and experienced personnel. We have developed a rapid method to detect Shiga toxin and SLT-I (Verotoxin 1) based on their binding to globotriosyl ceramide (Gb). Bound toxin was then detected by an enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. The direct detection of cytotoxins from pure culture plates and from a mixed bacterial culture was studied. Using polymyxin extraction (0.1 g/L, 30 min, 37°C) and Gb-based ELISA we detected toxin from reference strains 1 strain 60R (Shiga toxin) and 026: H11 strain H30 (SLT-I), and from clinical isolates of O157: H7 and O26: H11 (both SLT-I) from 11 patients with diarrhoea, haemorrhagic colitis or haemolytic uraemic syndrome. Toxin production by these strains was confirmed by a radiolabelled HeLa cell assay and the structural genes were detected by DNA hybridisation. The Gb-based ELISA could detect SLT-I in extracts of a mixed culture even when the toxin-positive strains represented only 1% of the mixture. No cross-reactivity was found with bacteria that produce other cytotoxins, such as other and and spp.

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1990-08-01
2022-01-28
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References

  1. Karmali MA. Infection by verocytotoxin-producing Esche richia coli. Clin Microbiol Rev 1989; 2:15–38
    [Google Scholar]
  2. Riley LW, Remis RS, Helgerson SD et al. Hemorrhagic colitis associated with rare Escherichia coli serotype. N Engl J Med 1983; 308:681–685
    [Google Scholar]
  3. Scotland SM, Willshaw GA, Smith HR, Rowe B. Properties of strains of Escherichia coli belonging to serogroup 0157 with special reference to production of Verocytotoxin VT1 and cytotoxin VT2. Epidemiol Infect 1987; 99:613–624
    [Google Scholar]
  4. Karmali MA, Petrie M, Lim C, Flemming PC, Arbus GS, Lior H. The association between idiopathic hemolytic uremic syndrome and infection by Verotoxin-producing Escherichia coli. J Infect Dis 1985; 151:775–782
    [Google Scholar]
  5. Neill MA, Tarr PI, Clausen CR, Christie DL, Hickman RO. Escherichia coli 0157 :H7 as the predominant pathogen associated with the hemolytic uremic syndrome: a prospective study in the Pacific Northwest. Pediatrics 1987; 80:37–40
    [Google Scholar]
  6. Cleary TG. Cytotoxin-producing Escherichia coli and hemolytic uremic syndrome. Pediatr Clin North Am 1988; 35:485–501
    [Google Scholar]
  7. Bartlett A V, Prado D, Cleary TG, Pickering LK. Production of Shiga toxin and other cytotoxins by serogroups of Shigella. J Infect Dis 1986; 154:996–1002
    [Google Scholar]
  8. Newland JW, Neill RJ. DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages. J Clin Microbiol 1988; 26:1292–1297
    [Google Scholar]
  9. Konowalchuk J, Speirs JI, Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun 1977; 18:775–779
    [Google Scholar]
  10. Strockbine NA, Marques LRM, Newland JW, Smith HW, Holmes RK, O’Brien AD. Two toxin-converting phages from Escherichia coli 0157 :H7 strain 933 encode antigenically distinct toxins with similar biologic activities. Infect Immun 1986; 53:135–140
    [Google Scholar]
  11. Strockbine NA, Marques LRM, Holmes RK, O’Brien AD. Characterization of monoclonal antibodies against Shiga-like toxin from Escherichia coli. Infect Immun 1985; 50:695–700
    [Google Scholar]
  12. Perera LP, Marques LRM, O’Brien AD. Isolation and characterization of monoclonal antibodies to Shiga-like toxin II of enterohemorrhagic Escherichia coli and use of the monoclonal antibodies in a colony enzyme-linked immunosorbent assay. J Clin Microbiol 1988; 26:2127–2131
    [Google Scholar]
  13. Jackson MP, Newland JW, Holmes RK, O’Brien AD. Nucleotide-sequence analysis of the structural genes for Shiga-like toxin I encoded by bacteriophage 933J from Escherichia coli. Microb Pathog 1987; 2:147–153
    [Google Scholar]
  14. Fuchs G, Mobassaleh M, Donohue-Rolfe A, Montgomery RK, Grand RJ, Keusch GT. Pathogenesis of shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for shigella toxin. Infect Immun 1986; 53:372–377
    [Google Scholar]
  15. Jacewicz M, Clausen H, Nudelman E, Donohue-Rolfe A, Kensch GT. Pathogenesis of shigella diarrhea. XI. Isolation of a shigella toxin-binding glycolipid from rabbit jejunum and HeLa cells and its identification as globotriaosylceramide. J Exp Med 1986; 163:1391–1404
    [Google Scholar]
  16. Lindberg AA, Brown JE, Stromberg N et al. Identification of the carbohydrate receptor for Shiga toxin produced by Shigella dysenteriae type 1. J Biol Chem 1987; 262:1779–1785
    [Google Scholar]
  17. Lingwood CA, Law H, Richardson SE et al. Glycolipid binding of purified and recombinant Escherichia coli produced verotoxin in vitro. J Biol Chem 1987; 262:8834–8839
    [Google Scholar]
  18. O’Brien AD, Holmes RK. Shiga and Shiga-like toxins. Microbiol Rev 1987; 51:206–220
    [Google Scholar]
  19. Weinstein DL, Jackson MP, Samuel JE, Holmes RK, O’Brien AD. Cloning and sequencing of a Shiga-like toxin type II variant from an Escherichia coli strain responsible for edema disease of swine. J Bacteriol 1988; 170:4223–4230
    [Google Scholar]
  20. Oku Y, Yutsudo T, Hirayama T, O’Brien AD, Takeda Y. Purification and some properties of a Vero toxin from a human strain of Escherichia coli that is immunologically related to Shiga-like toxin II (VT2). Microb Pathog 1989; 6:113–122
    [Google Scholar]
  21. Karch H, Meyer T. Evaluation of oligonucleotide probes for identification of Shiga-like-toxin-producing Escherichia coli. J Clin Microbiol 1989; 27:1180–1186
    [Google Scholar]
  22. Donohue-Rolfe A, Kelley MA, Bennish M, Keusch GT. Enzyme-linked immunosorbent assay for shigella toxin.. J Clin Microbiol 1986; 24:65–68
    [Google Scholar]
  23. Kongmuang U, Honda T, Miwatani T. Enzyme-linked immunosorbent assay to detect Shiga toxin of Shigella dysenteriae and related toxins. J Clin Microbiol 1987; 25:115–118
    [Google Scholar]
  24. Downes FP, Green JH, Greene K, Stockbrine W, Wells JG, Wachomulh IK. Development and evaluation of enzyme-linked immunosorbent assays for detection of Shiga-like toxin I and Shiga-like toxin II. J Clin Microbiol 1989; 27:1292–1297
    [Google Scholar]
  25. Ashkenazi S, Cleary TG. Rapid method to detect Shiga toxin and Shiga-like toxin I based on binding to globotriosyl ceramide (Gb3), their natural receptor. J Clin Microbiol 1989; 27:1145–1150
    [Google Scholar]
  26. Griffin DE, Gemski P. Release of Shiga toxin from Shigella dysenteriae 1 by polymyxin B. Infect Immun 1983; 40:425–428
    [Google Scholar]
  27. Karmali MA, Petrie M, Lim C, Cheung R, Arbus GS. Sensitive methods for detecting low numbers of verotoxin-producing Escherichia coli in mixed cultures by use of colony sweeps and polymyxin extraction of verotoxin. J Clin Microbiol 1985; 22:614–619
    [Google Scholar]
  28. Scotland SM, Rowe B, Smith HR, Willshaw GA, Gross RJ. Verocytotoxin-producing strains of Escherichia coli from children with haemolytic uraemic syndrome and their detection by specific DNA probes. J Med Microbiol 1988; 25:237–243
    [Google Scholar]
  29. Cleary TG, Mathewson JJ, Faris E, Pickering LK. Shiga-like cytotoxin production by enteropathogenic Escherichia coli serogroups. Infect Immun 1985; 47:335–337
    [Google Scholar]
  30. Prado D, Cleary TC, Pickering LK et al. The relation between production of cytotoxin and clinical features of shigellosis. J Infect Dis 1986; 154:149–155
    [Google Scholar]
  31. Ashkenazi S, Cleary TG, Murray BE, Wanger A, Pickering LK. Quantitative analysis and partial characterization of cytotoxin production by Salmonella strains. Infect Immun 1988; 56:3089–3094
    [Google Scholar]
  32. Kindschuh M, Pickering LK, Cleary TG, Ruis-Palacios G. Clinical and biochemical significance of toxin production by Aeromonas hydrophila. J Clin Microbiol 1987; 25:916–921
    [Google Scholar]
  33. Mathewson JJ, Cravioto A. HEp-2 adherence as an assay for virulence among diarrheagenic Escherichia coli. J Infect Dis 1989; 159:1057–1060
    [Google Scholar]
  34. Newland JW, Strockbine NA, Miller SF, O’Brien AD, Holmes RK. Cloning of Shiga-like toxin structural genes from a toxin converting phage of Escherichia coli. Science 1985; 230:179–181
    [Google Scholar]
  35. Maas R. An improved colony hybridization method with significantly increased sensitivity for detection of single genes. Plasmid 1983; 10:296–298
    [Google Scholar]
  36. Perry MB, Bundle DR, Gidney MAJ, Lior H. Identification of Escherichia coli serotype 0157 strains by using a monoclonal antibody. J Clin Microbiol 1988; 26:2391–2394
    [Google Scholar]
  37. March SB, Ratnam S. Latex agglutination test for detection of Escherichia coli serotype 0157. J Clin Microbiol 1989; 27:1675–1677
    [Google Scholar]
  38. Basta M, Karmali M, Lingwood C. Sensitive receptor-specific enzyme-linked immunosorbent assay for Escherichia coli verotoxin. J Clin Microbiol 1989; 27:1617–1622
    [Google Scholar]
  39. Waddell T, Head S, Petrie M, Cohen A, Lingwood C. Globotriosyl ceramide is specifically recognized by the Escherichia coli Vero cytotoxin 2. Biochem Biophys Res Commun 1988; 152:674–679
    [Google Scholar]
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