1887

Abstract

Summary

An enzyme-linked immunosorbent assay (ELISA) consisting of a double sandwich technique with rabbit and sheep antibodies, was developed for the detection of cytotoxic necrotising factor (CNF) in extracts of strains. The assay was evaluated by comparison with the results obtained with an assay based on toxicity for HeLa cell cultures. In a study of extracts of 27 CNF and 45 CNF strains obtained by ultrasonic disintegration, no false positive and only three false negative results were recorded; the latter were obtained with strains that produced less CNF than any of the others examined. Frozen-thawed extracts contained about four times more CNF cytotoxic activity than extracts prepared ultrasonically; the testing of 54 CNF and 68 CNF frozen-thawed extracts resulted in no false positive and only one false negative response Whichever type of extract was used, no significant cross-reaction was observed with heat-labile (LT) or heat stable (ST) enterotoxin, verotoxin (VT1, VT2), haemolysin, or Vir cytotoxin.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-32-2-73
1990-06-01
2022-01-27
Loading full text...

Full text loading...

/deliver/fulltext/jmm/32/2/medmicro-32-2-73.html?itemId=/content/journal/jmm/10.1099/00222615-32-2-73&mimeType=html&fmt=ahah

References

  1. Caprioli A., Falbo V., Roda L G., Ruggeri F M., Zona C. Partial purification and characterization of an Escherichia coli toxic factor that induces morphological cell alterations. Infect Immun 1983; 39:1300–1306
    [Google Scholar]
  2. Caprioli A., Donelli G., Falbo L. et al. A cell division-active protein from Escherichia coli. Biochem Biophys Res Commun 1984; 118:587–593
    [Google Scholar]
  3. De Rycke J., Phan-Thanh L., Bernard S. Immunochemical identification and biological characterization of cyto-toxic necrotizing factor of Escherichia coli. J Clin Microbiol 1989; 27:983–988
    [Google Scholar]
  4. Caprioli A., Falbo V., Ruggeri F M. et al. Cytotoxicnecrotising factor production by hemolytic strains of Escherichia coli causing extra-intestinal infections. J Clin Microbiol 1987; 25:146–149
    [Google Scholar]
  5. Alonso P., Blanco J., Blanco M., Gonzalez E A. Frequent production of toxins by Escherichia coli strains isolated from human urinary tract infections: relation with haemagglutination. FEMS Microbiol Lett 1987; 48:391–396
    [Google Scholar]
  6. Gonzalez E A., Blanco J. Production of cytotoxin VT in enteropathogenic and non-enteropathogenic Escherichia coli strains of porcine origin. FEMS Microbiol Lett 1985; 26:127–130
    [Google Scholar]
  7. De Rycke J., Guillot J F., Boivin R. Cytotoxins in non enterotoxigenic strains of Escherichia coli isolated from feces of diarrheic calves. Vet Microbiol 1987; 15:137–150
    [Google Scholar]
  8. Blanco J., Gonzalez E A., Garcia S., Blanco M., Regueiro B., Bemardez I. Production of toxins by Escherichia coli strains isolated from calves with diarrhoea in Galicia (North-western Spain). Vet Microbiol 1988; 18:297–311
    [Google Scholar]
  9. McLaren I., Wray C. Another animal Escherichia coli cytopathic factor. Vet Rec 1986; 119:576–577
    [Google Scholar]
  10. Smith H W. A search for transmissible pathogenic characters in invasive strains of Escherichia coli: the discovery of a plasmid-mediated toxin and a plasmid-controlled lethal character closely associated, or identical, with colicine V. J Gen Microbiol 1974; 83:95–111
    [Google Scholar]
  11. Oswald E., De Rycke J., Guillot J F., Boivin R. Cytotoxic effect of multinucleation in HeLa cell cultures associated with the presence of Vir plasmid in Escherichia coli strains. FEMS Microbiol Lett 1989; 58:95–100
    [Google Scholar]
  12. Bradford M M. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254
    [Google Scholar]
  13. McKinney M M., Parkinson A. A simple, non-chromatographic procedure to purify immunoglobulins from serum and ascites fluid. J Immunol Methods 1987; 96:271–278
    [Google Scholar]
  14. Guesdon J L., Avrameas S. Polyacrylamide-agarose beads for the preparation of effective immunoadsorbents. J Immunol Methods 1976; 11:129–133
    [Google Scholar]
  15. Tijssen P. Enzyme Immunoassays. In: Richard B., Kohler M D. eds Antigen detection to diagnose bacterial infections, vol. 1, Methodology. Boca Raton, CRC Press; 198699–135
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-32-2-73
Loading
/content/journal/jmm/10.1099/00222615-32-2-73
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