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. 1989 Feb;27(2):331–334. doi: 10.1128/jcm.27.2.331-334.1989

Examination of colonies and stool blots for detection of enteropathogens by DNA hybridization with eight DNA probes.

P Echeverria 1, D N Taylor 1, J Seriwatana 1, J E Brown 1, U Lexomboon 1
PMCID: PMC267302  PMID: 2563385

Abstract

We compared three methods for detecting enteropathogens in 416 children with diarrhea: (i) examination of 10 lactose-fermenting and all non-lactose-fermenting Escherichia coli (colony blots); (ii) examination of 300 colonies (replicate blots); and (iii) determination of the total bacterial growth of stools (stool blots). All specimens were spotted onto Whatman 541 filters and hybridized with specific radiolabeled DNA probes. Enterotoxigenic E. coli was detected in 38 patients by examining colony blots, in 52 patients by examining replicate blots, and in 45 patients by examining stool blots. Enteropathogenic E. coli adhesin factor was detected in 12 patients by colony blots, in 25 patients by replicate blots, and in 16 patients by stool blots. E. coli that hybridized with the enterohemorrhagic E. coli probe was detected in 2 patients by colony blots, in 11 patients by replicate blots, and in 0 patients by stool blots. Shiga-like toxin-producing E. coli was detected in 0 patients by colony blots, in 12 patients by replicate blots, and in 0 patients by stool blots. Shigella spp. were identified by standard bacteriological methods in 82 patients, and enteroinvasive E. coli was identified by colony blots in 11 patients (total, 93), by replicate blots in 56 patients, and by stool blots in 35 patients. Of 82 culture-confirmed Shigella infections, 45 were identified by examining replicate blots with the 17-kilobase-pair probe and 36 were identified by examination with the Ipa probe (P less than 0.05). Examining replicate blots with specific probes identified more enterotoxigenic E.coli (P < 0.005), enteropathogenic E.coli adhesion factor-producing E.coli (P < 0.001), and Shiga-like toxin-producing E.coli (P < 0.005) infections than examining colony blots. More Shigella and enteroinvasive E.coli infections were identified by standard bacteriological methods and examining colony blots with a specific probe than by examining replicate and stool blots (P < 0.001).

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Selected References

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  1. Buysse J. M., Stover C. K., Oaks E. V., Venkatesan M., Kopecko D. J. Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping. J Bacteriol. 1987 Jun;169(6):2561–2569. doi: 10.1128/jb.169.6.2561-2569.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chatkaeomorakot A., Echeverria P., Taylor D. N., Bettelheim K. A., Blacklow N. R., Sethabutr O., Seriwatana J., Kaper J. HeLa cell-adherent Escherichia coli in children with diarrhea in Thailand. J Infect Dis. 1987 Oct;156(4):669–672. doi: 10.1093/infdis/156.4.669. [DOI] [PubMed] [Google Scholar]
  3. 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]
  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. Gross R. J., Rowe B. Escherichia coli diarrhoea. J Hyg (Lond) 1985 Dec;95(3):531–550. doi: 10.1017/s0022172400060666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Karch H., Heesemann J., Laufs R., O'Brien A. D., Tacket C. O., Levine M. M. A plasmid of enterohemorrhagic Escherichia coli O157:H7 is required for expression of a new fimbrial antigen and for adhesion to epithelial cells. Infect Immun. 1987 Feb;55(2):455–461. doi: 10.1128/iai.55.2.455-461.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Konowalchuk J., Speirs J. I., Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977 Dec;18(3):775–779. doi: 10.1128/iai.18.3.775-779.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Levine M. M., Xu J. G., Kaper J. B., Lior H., Prado V., Tall B., Nataro J., Karch H., Wachsmuth K. A DNA probe to identify enterohemorrhagic Escherichia coli of O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis. 1987 Jul;156(1):175–182. doi: 10.1093/infdis/156.1.175. [DOI] [PubMed] [Google Scholar]
  9. Maas R. An improved colony hybridization method with significantly increased sensitivity for detection of single genes. Plasmid. 1983 Nov;10(3):296–298. doi: 10.1016/0147-619x(83)90045-8. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Nataro J. P., Baldini M. M., Kaper J. B., Black R. E., Bravo N., Levine M. M. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. doi: 10.1093/infdis/152.3.560. [DOI] [PubMed] [Google Scholar]
  12. Nataro J. P., Scaletsky I. C., Kaper J. B., Levine M. M., Trabulsi L. R. Plasmid-mediated factors conferring diffuse and localized adherence of enteropathogenic Escherichia coli. Infect Immun. 1985 May;48(2):378–383. doi: 10.1128/iai.48.2.378-383.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture. Infect Immun. 1975 Feb;11(2):334–336. doi: 10.1128/iai.11.2.334-336.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Scaletsky I. C., Silva M. L., Trabulsi L. R. Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells. Infect Immun. 1984 Aug;45(2):534–536. doi: 10.1128/iai.45.2.534-536.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Seriwatana J., Echeverria P., Taylor D. N., Rasrinaul L., Brown J. E., Peiris J. S., Clayton C. L. Type II heat-labile enterotoxin-producing Escherichia coli isolated from animals and humans. Infect Immun. 1988 May;56(5):1158–1161. doi: 10.1128/iai.56.5.1158-1161.1988. [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. Smith H. R., Rowe B., Gross R. J., Fry N. K., Scotland S. M. Haemorrhagic colitis and Vero-cytotoxin-producing Escherichia coli in England and Wales. Lancet. 1987 May 9;1(8541):1062–1065. doi: 10.1016/s0140-6736(87)90485-5. [DOI] [PubMed] [Google Scholar]
  18. So M., Dallas W. S., Falkow S. Characterization of an Escherichia coli plasmid encoding for synthesis of heat-labile toxin: molecular cloning of the toxin determinant. Infect Immun. 1978 Aug;21(2):405–411. doi: 10.1128/iai.21.2.405-411.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Taylor D. N., Echeverria P., Pál T., Sethabutr O., Saiborisuth S., Sricharmorn S., Rowe B., Cross J. The role of Shigella spp., enteroinvasive Escherichia coli, and other enteropathogens as causes of childhood dysentery in Thailand. J Infect Dis. 1986 Jun;153(6):1132–1138. doi: 10.1093/infdis/153.6.1132. [DOI] [PubMed] [Google Scholar]
  20. Taylor D. N., Echeverria P., Sethabutr O., Pitarangsi C., Leksomboon U., Blacklow N. R., Rowe B., Gross R., Cross J. Clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli infections detected by DNA hybridization. J Clin Microbiol. 1988 Jul;26(7):1362–1366. doi: 10.1128/jcm.26.7.1362-1366.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Venkatesan M., Buysse J. M., Vandendries E., Kopecko D. J. Development and testing of invasion-associated DNA probes for detection of Shigella spp. and enteroinvasive Escherichia coli. J Clin Microbiol. 1988 Feb;26(2):261–266. doi: 10.1128/jcm.26.2.261-266.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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