Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 May;32(5):1246–1248. doi: 10.1128/jcm.32.5.1246-1248.1994

Direct detection of Vibrio cholerae in stool samples.

P Varela 1, G D Pollevick 1, M Rivas 1, I Chinen 1, N Binsztein 1, A C Frasch 1, R A Ugalde 1
PMCID: PMC263658  PMID: 8051251

Abstract

A direct method to detect Vibrio cholerae in stool samples was developed by using a PCR procedure that did not require a DNA purification step. Dilution (1/100) of stool samples prevented inhibition of the reaction by contaminants, and two consecutive PCRs, the second one with a nested primer, achieved the desired sensitivity. Comparison of the results obtained from stool swab samples processed by the two-step PCR and by an enzyme-linked immunosorbent assay using GM1 as the capture molecule showed that the former is more sensitive and gave positive results even when V. cholerae was not culturable or dead.

Full text

PDF
1246

Images in this article

1247Image
on p.1247
1247Image
on p.1247

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Craig J. P., Yamamoto K., Takeda Y., Miwatani T. Production of cholera-like enterotoxin by a Vibrio cholerae non-O1 strain isolated from the environment. Infect Immun. 1981 Oct;34(1):90–97. doi: 10.1128/iai.34.1.90-97.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fields P. I., Popovic T., Wachsmuth K., Olsvik O. Use of polymerase chain reaction for detection of toxigenic Vibrio cholerae O1 strains from the Latin American cholera epidemic. J Clin Microbiol. 1992 Aug;30(8):2118–2121. doi: 10.1128/jcm.30.8.2118-2121.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Guidolin A., Manning P. A. Genetics of Vibrio cholerae and its bacteriophages. Microbiol Rev. 1987 Jun;51(2):285–298. doi: 10.1128/mr.51.2.285-298.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Iwanaga M., Yamamoto K. New medium for the production of cholera toxin by Vibrio cholerae O1 biotype El Tor. J Clin Microbiol. 1985 Sep;22(3):405–408. doi: 10.1128/jcm.22.3.405-408.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kaper J. B., Moseley S. L., Falkow S. Molecular characterization of environmental and nontoxigenic strains of Vibrio cholerae. Infect Immun. 1981 May;32(2):661–667. doi: 10.1128/iai.32.2.661-667.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kato N., Ou C. Y., Kato H., Bartley S. L., Luo C. C., Killgore G. E., Ueno K. Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction. J Infect Dis. 1993 Feb;167(2):455–458. doi: 10.1093/infdis/167.2.455. [DOI] [PubMed] [Google Scholar]
  7. Koch W. H., Payne W. L., Wentz B. A., Cebula T. A. Rapid polymerase chain reaction method for detection of Vibrio cholerae in foods. Appl Environ Microbiol. 1993 Feb;59(2):556–560. doi: 10.1128/aem.59.2.556-560.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mekalanos J. J., Swartz D. J., Pearson G. D., Harford N., Groyne F., de Wilde M. Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development. Nature. 1983 Dec 8;306(5943):551–557. doi: 10.1038/306551a0. [DOI] [PubMed] [Google Scholar]
  9. Rivas M., Binsztein N., López Moral L., Cinto R. Produccion de enterotoxina termolábil por cepas de Escherichia coli aisladas en Argentina. Rev Argent Microbiol. 1987 Jul-Sep;19(3):91–100. [PubMed] [Google Scholar]
  10. Shirai H., Nishibuchi M., Ramamurthy T., Bhattacharya S. K., Pal S. C., Takeda Y. Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae. J Clin Microbiol. 1991 Nov;29(11):2517–2521. doi: 10.1128/jcm.29.11.2517-2521.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Varela P., Rivas M., Binsztein N., Cremona M. L., Herrmann P., Burrone O., Ugalde R. A., Frasch A. C. Identification of toxigenic Vibrio cholerae from the Argentine outbreak by PCR for ctx A1 and ctx A2-B. FEBS Lett. 1993 Jan 2;315(1):74–76. doi: 10.1016/0014-5793(93)81136-n. [DOI] [PubMed] [Google Scholar]
  12. Wilde J., Eiden J., Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol. 1990 Jun;28(6):1300–1307. doi: 10.1128/jcm.28.6.1300-1307.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Yoh M., Miyagi K., Matsumoto Y., Hayashi K., Takarada Y., Yamamoto K., Honda T. Development of an enzyme-labeled oligonucleotide probe for the cholera toxin gene. J Clin Microbiol. 1993 May;31(5):1312–1314. doi: 10.1128/jcm.31.5.1312-1314.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES