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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1988 Jan;26(1):92–95. doi: 10.1128/jcm.26.1.92-95.1988

Alkaline phosphatase-conjugated oligonucleotide probes for enterotoxigenic Escherichia coli in travelers to South America and West Africa.

J J Oprandy 1, S A Thornton 1, C H Gardiner 1, D Burr 1, R Batchelor 1, A L Bourgeois 1
PMCID: PMC266198  PMID: 3277993

Abstract

Several studies have demonstrated the usefulness of 32P-labeled recombinant DNA probes for identifying enterotoxigenic Escherichia coli (ETEC). The use of radioisotopes and X-ray development, however, severely handicaps the utility of DNA probes in most clinical laboratories. In this study, enzyme-labeled oligonucleotide probes for ETEC LT (heat-labile toxin) and ST (heat-stable toxin) genes were compared with the standard Y1 adrenal cell and suckling mouse assays for their ability to identify ETEC in a population of American adults experiencing acute episodes of diarrhea in South America and West Africa. The LT probe hybridized with 12% (64 of 529) of the E. coli colonies tested, whereas 11% (57 of 529) were positive by Y1 adrenal cell assay. DNA from 9% (47 of 529) of the E. coli colonies tested hybridized with the ST probe, whereas only 5% (28 of 529) produced ST as measured by the suckling mouse bioassay. For the patient samples tested, correlation between probe and bioassay for LT was 97%, or three discrepancies in 111 patients tested. Overall concordance of the ST probe and bioassay was 95%, or five discrepancies in 111 patients. Enzyme-labeled oligonucleotide probes represent a major advance in the diagnosis of ETEC-associated diarrheal disease and may be used in laboratories with minimal equipment.

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Selected References

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

  1. Bialkowska-Hobrzanska H. Detection of enterotoxigenic Escherichia coli by dot blot hybridization with biotinylated DNA probes. J Clin Microbiol. 1987 Feb;25(2):338–343. doi: 10.1128/jcm.25.2.338-343.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dean A. G., Ching Y. C., Williams R. G., Harden L. B. Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis. 1972 Apr;125(4):407–411. doi: 10.1093/infdis/125.4.407. [DOI] [PubMed] [Google Scholar]
  3. Donta S. T., Moon H. W., Whipp S. C. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science. 1974 Jan 25;183(4122):334–336. doi: 10.1126/science.183.4122.334. [DOI] [PubMed] [Google Scholar]
  4. Echeverria P., Seriwatana J., Chityothin O., Chaicumpa W., Tirapat C. Detection of enterotoxigenic Escherichia coli in water by filter hybridization with three enterotoxin gene probes. J Clin Microbiol. 1982 Dec;16(6):1086–1090. doi: 10.1128/jcm.16.6.1086-1090.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Echeverria P., Seriwatana J., Taylor D. N., Tirapat C., Chaicumpa W., Rowe B. Identification by DNA hybridization of enterotoxigenic Escherichia coli in a longitudinal study of villages in Thailand. J Infect Dis. 1985 Jan;151(1):124–130. doi: 10.1093/infdis/151.1.124. [DOI] [PubMed] [Google Scholar]
  6. Echeverria P., Taylor D. N., Seriwatana J., Chatkaeomorakot A., Khungvalert V., Sakuldaipeara T., Smith R. D. A comparative study of enterotoxin gene probes and tests for toxin production to detect enterotoxigenic Escherichia coli. J Infect Dis. 1986 Feb;153(2):255–260. doi: 10.1093/infdis/153.2.255. [DOI] [PubMed] [Google Scholar]
  7. Echeverria P., Taylor D. N., Seriwatana J., Moe C. Comparative study of synthetic oligonucleotide and cloned polynucleotide enterotoxin gene probes to identify enterotoxigenic Escherichia coli. J Clin Microbiol. 1987 Jan;25(1):106–109. doi: 10.1128/jcm.25.1.106-109.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans D. G., Olarte J., DuPont H. L., Evans D. J., Jr, Galindo E., Portnoy B. L., Conklin R. H. Enteropathogens associated with pediatric diarrhea in Mexico City. J Pediatr. 1977 Jul;91(1):65–68. doi: 10.1016/s0022-3476(77)80446-0. [DOI] [PubMed] [Google Scholar]
  9. Guerrant R. L., Kirchhoff L. V., Shields D. S., Nations M. K., Leslie J., de Sousa M. A., Araujo J. G., Correia L. L., Sauer K. T., McClelland K. E. Prospective study of diarrheal illnesses in northeastern Brazil: patterns of disease, nutritional impact, etiologies, and risk factors. J Infect Dis. 1983 Dec;148(6):986–997. doi: 10.1093/infdis/148.6.986. [DOI] [PubMed] [Google Scholar]
  10. Lockwood D. E., Robertson D. C. Development of a competitive enzyme-linked immunosorbent assay (ELISA) for Escherichia coli heat-stable enterotoxin (STa). J Immunol Methods. 1984 Dec 31;75(2):295–307. doi: 10.1016/0022-1759(84)90113-3. [DOI] [PubMed] [Google Scholar]
  11. Moseley S. L., Echeverria P., Seriwatana J., Tirapat C., Chaicumpa W., Sakuldaipeara T., Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. doi: 10.1093/infdis/145.6.863. [DOI] [PubMed] [Google Scholar]
  12. Moseley S. L., Hardy J. W., Hug M. I., Echeverria P., Falkow S. Isolation and nucleotide sequence determination of a gene encoding a heat-stable enterotoxin of Escherichia coli. Infect Immun. 1983 Mar;39(3):1167–1174. doi: 10.1128/iai.39.3.1167-1174.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Murray B. E., Mathewson J. J., DuPont H. L., Hill W. E. Utility of oligodeoxyribonucleotide probes for detecting enterotoxigenic Escherichia coli. J Infect Dis. 1987 Apr;155(4):809–811. doi: 10.1093/infdis/155.4.809. [DOI] [PubMed] [Google Scholar]
  14. Seriwatana J., Echeverria P., Escamilla J., Glass R., Huq I., Rockhill R., Stoll B. J. Identification of enterotoxigenic Escherichia coli in patients with diarrhea in Asia with three enterotoxin gene probes. Infect Immun. 1983 Oct;42(1):152–155. doi: 10.1128/iai.42.1.152-155.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Seriwatana J., Echeverria P., Taylor D. N., Sakuldaipeara T., Changchawalit S., Chivoratanond O. Identification of enterotoxigenic Escherichia coli with synthetic alkaline phosphatase-conjugated oligonucleotide DNA probes. J Clin Microbiol. 1987 Aug;25(8):1438–1441. doi: 10.1128/jcm.25.8.1438-1441.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sethabutr O., Hanchalay S., Echeverria P., Taylor D. N., Leksomboon U. A non-radioactive DNA probe to identify Shigella and enteroinvasive Escherichia coli in stools of children with diarrhoea. Lancet. 1985 Nov 16;2(8464):1095–1097. doi: 10.1016/s0140-6736(85)90687-7. [DOI] [PubMed] [Google Scholar]
  17. Thompson M. R., Jordan R. L., Luttrell M. A., Brandwein H., Kaper J. B., Levine M. M., Giannella R. A. Blinded, two-laboratory comparative analysis of Escherichia coli heat-stable enterotoxin production by using monoclonal antibody enzyme-linked immunosorbent assay, radioimmunoassay, suckling mouse assay, and gene probes. J Clin Microbiol. 1986 Nov;24(5):753–758. doi: 10.1128/jcm.24.5.753-758.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yamamoto T., Tamura T., Yokota T. Primary structure of heat-labile enterotoxin produced by Escherichia coli pathogenic for humans. J Biol Chem. 1984 Apr 25;259(8):5037–5044. [PubMed] [Google Scholar]
  19. Zaki A. M., DuPont H. L., el Alamy M. A., Arafat R. R., Amin K., Awad M. M., Bassiouni L., Imam I. Z., el Malih G. S., el Marsafie A. The detection of enteropathogens in acute diarrhea in a family cohort population in rural Egypt. Am J Trop Med Hyg. 1986 Sep;35(5):1013–1022. doi: 10.4269/ajtmh.1986.35.1013. [DOI] [PubMed] [Google Scholar]

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