1887

Abstract

Summary

Techniques currently available to detect Shiga-like toxin (SLT)-producing lack sensitivity or require specialised equipment and facilities, and in some cases detect only strains belonging to serotype O157. We have used an ELISA technique, capable of detecting both SLTI and SLTII with crude P1 glycoprotein from hydatid cysts, in combination with enhancement of toxin production by culture with mitomycin C. Supernates of Tryptone Soya Broth cultures containing mitomycin C 200 ng/ml were tested for SLTII. For SLTI, cell lysates pre-treated with polymyxin B were tested. In tests with O157:H7 in mixed culture with strain C600 alone, or with C600, and , SLTI could be detected when the proportion of toxigenic organisms represented 1% of the mixture, and SLTII when the proportion was 0‡025%. When faecal samples with added O157:H7 were examined in this system, SLTII-producing strains were detected when they comprised < 0‡1% of the coliform population. This technique is a sensitive and specific assay for detecting low numbers of SLT-producing organisms in mixed culture such as occurs in cases of haemolytic uraemic syndrome and haemorrhagic colitis.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-36-3-198
1992-03-01
2022-01-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/36/3/medmicro-36-3-198.html?itemId=/content/journal/jmm/10.1099/00222615-36-3-198&mimeType=html&fmt=ahah

References

  1. Karmali M. A. Infection by verocytotoxin producing Escherichia coli. Clin Microbiol Rev 1989; 2:15–38
    [Google Scholar]
  2. Riley L. W. The epidemiologic, clinical, and microbiologic features of hemorrhagic colitis. Annu Rev Microbiol 1987; 41:383–407
    [Google Scholar]
  3. Milford D. V., Taylor C. M. New insights into the haemolytic uraemic syndromes. Arch Dis Child 1990; 65:713–715
    [Google Scholar]
  4. O’Brien A. D., Holmes R. K. Shiga and Shiga-like toxins. Microbiol Rev 1987; 51:206–220
    [Google Scholar]
  5. Strockbine N. A., Marques L. R. M., Newland J. W., Williams Smith H., Holmes R. K., O’Brien A. D. Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode anti-genically distinct toxins with similar biologic activities. Infect Immun 1986; 53:135–140
    [Google Scholar]
  6. Mobassaleh M., Donohue-Rolfe A., Jacewicz M., Grand R. J., Keusch G. T. Pathogenesis of Shigella diarrhea: evidence for a developmentally regulated glycolipid receptor for shigella toxin involved in the fluid secretory response of rabbit small intestine. J Infect Dis 1988; 157:1023–1031
    [Google Scholar]
  7. Lior H., Borczyk A. A. False positive identification of Escherichia coli O157. Lancet 1987; 1:333
    [Google Scholar]
  8. Kleanthous H., Smith H. R., Scotland S. M. Haemolytic uraemic syndromes in the British Isles, 1985-8: association with Verocytotoxin producing Escherichia coli. Part 2: microbiological aspects. Arch Dis Child 1990; 65:722–727
    [Google Scholar]
  9. Bettelheim K. A., Brown J. E., Lolekha S., Echeverria P. Serotypes of Escherichia coli that hybridized with DNA probes for genes encoding Shiga-like toxin I, Shiga-like toxin II, and serogroup O157 enterohemorrhagic E. coli fimbriae isolated from adults with diarrhea in Thailand. J Clin Microbiol 1990; 28:293–295
    [Google Scholar]
  10. Scotland S. M., Rowe B., Smith H. R., Willshaw G. A., Gross R. J. Vero cytotoxin-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]
  11. Newland J. W., Neill R. J. DNA probes for Shiga-like toxin I and II and for toxin-converting bacteriophages. J Clin Microbiol 1988; 26:1292–1297
    [Google Scholar]
  12. Karch H., Meyer T. Single primer pair for amplifying segments of distinct Shiga-like-toxin genes by polymerase chain reaction. J Clin Microbiol 1989; 27:2751–2757
    [Google Scholar]
  13. Pollard D. R., Johnson W. M., Lior H., Tyler S. D., Rozee K. R. Rapid and specific detection of verocytotoxin genes in Escherichia coli by the polymerase chain reaction. J Clin Microbiol 1990; 28:540–545
    [Google Scholar]
  14. Downes F., Green J. H., Greene K., Strockbine N., Wells J. G., Wachsmuth I. K. 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]
  15. Ashkenazi S., Cleary T. G. 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]
  16. Basta M., Karmali M., Lingwood C. Sensitive receptor-specified enzyme-linked immunosorbent assay for Escherichia coli verocytotoxin. J Clin Microbiol 1989; 27:1617–1622
    [Google Scholar]
  17. Donohue-Rolfe A., Acheson D. W. K., Kane A. V., Keusch G. T. Purification of Shiga toxin and Shiga-like toxins I and II by receptor analog affinity chromatography with immobilized P1 glycoprotein and the production of cross-reactive monoclonal antibodies. Infect Immun 1989; 57:3888–3893
    [Google Scholar]
  18. Acheson D. W. K., Keusch G. T., Lightowlers M., Donohue-Rolfe A. Enzyme-linked immunosorbent assay for Shiga toxin and Shiga-like toxin II using P, glycoprotein from hydatid cysts. J Infect Dis 1990; 161:134–137
    [Google Scholar]
  19. Head S. C., Petrie M., Richardson S., Roscoe M., Karmali M. A. Purification and characterization of verocytotoxin 2. FEMS Microbiol Lett 1988; 51:211–216
    [Google Scholar]
  20. Acheson D. W. K., Kane A. V., Keusch G. T., Donohue-Rolfe A. High yield purification and subunit characterisation of Shiga-like toxin II. Interscience Conference on Anti-microbial Agents and Chemotherapy 1989 Abstract no. 901 252
    [Google Scholar]
  21. Tarr P. I., Neill M. A., Clausen C. R., Watkins S. L., Christie D. L., Hickman R. O. Escherichia coli O157: H7 and the hemolytic uremic syndrome: importance of early cultures in establishing the etiology. J Infect Dis 1990; 162:553–556
    [Google Scholar]
  22. Ashkenazi S., Cleary T. G. A method for detecting Shiga toxin and Shiga-like toxin-I in pure and mixed culture. J Med Microbiol 1990; 32:255–261
    [Google Scholar]
  23. Wadolkowski E. A., Sung L. M., Burris J. A., Samuel J. E., O’Brien A. D. Acute renal tubular necrosis and death of mice orally infected with Escherichia coli strains that produce Shiga-like toxin type II. Infect Immun 1990; 58:3959–3965
    [Google Scholar]
  24. Tarr P. I., Neill M. A., Clausen C. R., Newland J. W., Neill R. J., Moseley S. L. Genotypic variation in pathogenic Escherichia coli O157:H7 isolated from patients in Washington, 1984-87. J Infect Dis 1989; 159:344–347
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-36-3-198
Loading
/content/journal/jmm/10.1099/00222615-36-3-198
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error