The use of gene probes, immunoassays and tissue culture for the detection of toxin in Vibrio cholerae non-O1

J Med Microbiol. 1994 Jan;40(1):31-6. doi: 10.1099/00222615-40-1-31.

Abstract

Vibrio cholerae non-O1 strains were screened for the presence of cholera enterotoxin (CT) genes by means of digoxigenin-labelled polynucleotide CTA and CTB probes. In-vitro production of CT was investigated by the Y1 mouse adrenal cell assay, enzyme-linked immunosorbent assay (ELISA) and a commercial, reversed passive latex agglutination (RPLA) kit. Only two (0.25%) of 790 strains tested gave positive results with the CTA and CTB probes. The production of other bacterial cytotoxin(s) made it impossible to use the characteristic cell-rounding effect on Y1 cells for the detection of CT. CT production by the probe-positive strains was confirmed by the immunoassays. Two hundred and fifty-two of the 788 probe-negative strains were tested by both cell assay and immunoassays. Of these, 90% produced cytotoxin(s) in the cell assay. In addition, 37% gave positive results in CT-ELISA, but negative results with LT-ELISA and VET-RPLA. These results indicate the presumed presence of a toxin in V. cholerae non-O1 that is able to bind GM1 and react with antisera to CT, but which is not identical to CT.

MeSH terms

  • Cell Line
  • Culture Media
  • DNA Probes
  • Enterotoxins / biosynthesis*
  • Enterotoxins / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Latex Fixation Tests
  • Nucleic Acid Hybridization
  • Vibrio cholerae / genetics
  • Vibrio cholerae / metabolism*

Substances

  • Culture Media
  • DNA Probes
  • Enterotoxins
  • stN protein, Vibrio cholerae