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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Oct;27(10):2272–2276. doi: 10.1128/jcm.27.10.2272-2276.1989

Detection with synthetic oligonucleotide probes of nucleotide sequence variations in the genes encoding enterotoxins of Escherichia coli.

M Nishibuchi 1, A Murakami 1, M Arita 1, H Jikuya 1, J Takano 1, T Honda 1, T Miwatani 1
PMCID: PMC267009  PMID: 2685027

Abstract

We examined variations in the genes encoding heat-stable enterotoxin (ST) and heat-labile enterotoxin (LT) in 88 strains of Escherichia coli isolated from individuals with traveler's diarrhea to find suitable sequences for use as oligonucleotide probes. Four oligonucleotide probes of the gene encoding ST of human origin (STIb or STh), one oligonucleotide probe of the gene encoding ST of porcine origin (STIa or STp), and three oligonucleotide probes of the gene encoding LT of human origin (LTIh) were used in DNA colony hybridization tests. In 15 of 22 strains possessing the STh gene and 28 of 42 strains producing LT, the sequences of all regions tested were identical to the published sequences. One region in the STh gene examined with a 18-mer probe was relatively well conserved and was shown to be closely associated with the enterotoxicity of the E. coli strains in suckling mice. This oligonucleotide, however, hybridized with strains of Vibrio cholerae O1, V. parahaemolyticus, and Yersinia enterocolitica that gave negative results in the suckling mouse assay.

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Selected References

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  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. Echeverria P., Seriwatana J., Leksomboon U., Tirapat C., Chaicumpa W., Rowe B. Identification by DNA hybridisation of enterotoxigenic Escherichia coli in homes of children with diarrhoea. Lancet. 1984 Jan 14;1(8368):63–66. doi: 10.1016/s0140-6736(84)90001-1. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. Hill W. E., Payne W. L., Zon G., Moseley S. L. Synthetic oligodeoxyribonucleotide probes for detecting heat-stable enterotoxin-producing Escherichia coli by DNA colony hybridization. Appl Environ Microbiol. 1985 Nov;50(5):1187–1191. doi: 10.1128/aem.50.5.1187-1191.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Honda T., Arita M., Takeda T., Yoh M., Miwatani T. Non-O1 Vibrio cholerae produces two newly identified toxins related to Vibrio parahaemolyticus haemolysin and Escherichia coli heat-stable enterotoxin. Lancet. 1985 Jul 20;2(8447):163–164. doi: 10.1016/s0140-6736(85)90277-6. [DOI] [PubMed] [Google Scholar]
  8. Honda T., Arita M., Takeda Y., Miwatani T. Further evaluation of the Biken test (modified Elek test) for detection of enterotoxigenic Escherichia coli producing heat-labile enterotoxin and application of the test to sampling of heat-stable enterotoxin. J Clin Microbiol. 1982 Jul;16(1):60–62. doi: 10.1128/jcm.16.1.60-62.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jablonski E., Moomaw E. W., Tullis R. H., Ruth J. L. Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes. Nucleic Acids Res. 1986 Aug 11;14(15):6115–6128. doi: 10.1093/nar/14.15.6115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kirii Y., Danbara H., Komase K., Arita H., Yoshikawa M. Detection of enterotoxigenic Escherichia coli by colony hybridization with biotinylated enterotoxin probes. J Clin Microbiol. 1987 Oct;25(10):1962–1965. doi: 10.1128/jcm.25.10.1962-1965.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Li P., Medon P. P., Skingle D. C., Lanser J. A., Symons R. H. Enzyme-linked synthetic oligonucleotide probes: non-radioactive detection of enterotoxigenic Escherichia coli in faecal specimens. Nucleic Acids Res. 1987 Jul 10;15(13):5275–5287. doi: 10.1093/nar/15.13.5275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Medon P. P., Lanser J. A., Monckton P. R., Li P., Symons R. H. Identification of enterotoxigenic Escherichia coli isolates with enzyme-labeled synthetic oligonucleotide probes. J Clin Microbiol. 1988 Oct;26(10):2173–2176. doi: 10.1128/jcm.26.10.2173-2176.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Nishibuchi M., Arita M., Honda T., Miwatani T. Evaluation of a nonisotopically labeled oligonucleotide probe to detect the heat-stable enterotoxin gene of Escherichia coli by the DNA colony hybridization test. J Clin Microbiol. 1988 Apr;26(4):784–786. doi: 10.1128/jcm.26.4.784-786.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nishibuchi M., Hill W. E., Zon G., Payne W. L., Kaper J. B. Synthetic oligodeoxyribonucleotide probes to detect Kanagawa phenomenon-positive Vibrio parahaemolyticus. J Clin Microbiol. 1986 Jun;23(6):1091–1095. doi: 10.1128/jcm.23.6.1091-1095.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Okamoto K., Inoue T., Shimizu K., Hara S., Miyama A. Further purification and characterization of heat-stable enterotoxin produced by Yersinia enterocolitica. Infect Immun. 1982 Mar;35(3):958–964. doi: 10.1128/iai.35.3.958-964.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Oprandy J. J., Thornton S. A., Gardiner C. H., Burr D., Batchelor R., Bourgeois A. L. Alkaline phosphatase-conjugated oligonucleotide probes for enterotoxigenic Escherichia coli in travelers to South America and West Africa. J Clin Microbiol. 1988 Jan;26(1):92–95. doi: 10.1128/jcm.26.1.92-95.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Sommerfelt H., Kalland K. H., Raj P., Moseley S. L., Bhan M. K., Bjorvatn B. Cloned polynucleotide and synthetic oligonucleotide probes used in colony hybridization are equally efficient in the identification of enterotoxigenic Escherichia coli. J Clin Microbiol. 1988 Nov;26(11):2275–2278. doi: 10.1128/jcm.26.11.2275-2278.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sommerfelt H., Svennerholm A. M., Kalland K. H., Haukanes B. I., Bjorvatn B. Comparative study of colony hybridization with synthetic oligonucleotide probes and enzyme-linked immunosorbent assay for identification of enterotoxigenic Escherichia coli. J Clin Microbiol. 1988 Mar;26(3):530–534. doi: 10.1128/jcm.26.3.530-534.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takao T., Shimonishi Y., Kobayashi M., Nishimura O., Arita M., Takeda T., Honda T., Miwatani T. Amino acid sequence of heat-stable enterotoxin produced by Vibrio cholerae non-01. FEBS Lett. 1985 Dec 2;193(2):250–254. doi: 10.1016/0014-5793(85)80163-0. [DOI] [PubMed] [Google Scholar]
  24. Takao T., Tominaga N., Yoshimura S., Shimonishi Y., Hara S., Inoue T., Miyama A. Isolation, primary structure and synthesis of heat-stable enterotoxin produced by Yersinia enterocolitica. Eur J Biochem. 1985 Oct 1;152(1):199–206. doi: 10.1111/j.1432-1033.1985.tb09183.x. [DOI] [PubMed] [Google Scholar]
  25. Wallace R. B., Shaffer J., Murphy R. F., Bonner J., Hirose T., Itakura K. Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch. Nucleic Acids Res. 1979 Aug 10;6(11):3543–3557. doi: 10.1093/nar/6.11.3543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yamamoto T., Nakazawa T., Miyata T., Kaji A., Yokota T. Evolution and structure of two ADP-ribosylation enterotoxins, Escherichia coli heat-labile toxin and cholera toxin. FEBS Lett. 1984 Apr 24;169(2):241–246. doi: 10.1016/0014-5793(84)80326-9. [DOI] [PubMed] [Google Scholar]
  27. Yoshimura S., Ikemura H., Watanabe H., Aimoto S., Shimonishi Y., Hara S., Takeda T., Miwatani T., Takeda Y. Essential structure for full enterotoxigenic activity of heat-stable enterotoxin produced by enterotoxigenic Escherichia coli. FEBS Lett. 1985 Feb 11;181(1):138–142. doi: 10.1016/0014-5793(85)81129-7. [DOI] [PubMed] [Google Scholar]

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