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. 1989 Dec;27(12):2717–2723. doi: 10.1128/jcm.27.12.2717-2723.1989

Species-specific cloned DNA probes for the identification of Campylobacter hyointestinalis.

C J Gebhart 1, G E Ward 1, M P Murtaugh 1
PMCID: PMC267116  PMID: 2592537

Abstract

Conventional microbiological methods for isolating and identifying Campylobacter species are laborious, tedious, and subjective. Because of the increasing importance of Campylobacter species in human and animal diseases and the recent emergence of many new species and atypical strains, we are developing chromosomal DNA probes for rapid and simple identification of Campylobacter species, especially those of veterinary importance. We report the cloning and characterization of chromosomal DNA fragments from Campylobacter hyointestinalis, an organism isolated from pigs with proliferative enteritis. To obtain C. hyointestinalis-specific probes, chromosomal DNA fragments from C. hyointestinalis were cloned into plasmid vector pGEM-3Z. Recombinant plasmids were screened for C. hyointestinalis-specific inserts by DNA hybridization, using chromosomal DNA from either C. hyointestinalis or C. fetus which had been 32P labeled. Recombinants which hybridized to C. hyointestinalis, but not C. fetus, DNA were 32P labeled and screened further for sensitivity and specificity. Three probes were identified that were species specific and capable of detecting 10(4) C. hyointestinalis organisms by bacterial spot blotting in 48 h. We anticipate that these probes will be useful for routine species identification and for epidemiological studies.

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Selected References

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

  1. Belland R. J., Trust T. J. Deoxyribonucleic acid sequence relatedness between thermophilic members of the genus Campylobacter. J Gen Microbiol. 1982 Nov;128(11):2515–2522. doi: 10.1099/00221287-128-11-2515. [DOI] [PubMed] [Google Scholar]
  2. Blaser M. J., Reller L. B. Campylobacter enteritis. N Engl J Med. 1981 Dec 10;305(24):1444–1452. doi: 10.1056/NEJM198112103052404. [DOI] [PubMed] [Google Scholar]
  3. Claus P., Moseley S. L., Falkow S. The use of gene-specific DNA probes for the identification of enteric pathogens. Prog Food Nutr Sci. 1983;7(3-4):139–142. [PubMed] [Google Scholar]
  4. Echeverria P., Seriwatana J., Sethabutr O., Taylor D. N. DNA hybridization in the diagnosis of bacterial diarrhea. Clin Lab Med. 1985 Sep;5(3):447–462. [PubMed] [Google Scholar]
  5. Edmonds P., Patton C. M., Griffin P. M., Barrett T. J., Schmid G. P., Baker C. N., Lambert M. A., Brenner D. J. Campylobacter hyointestinalis associated with human gastrointestinal disease in the United States. J Clin Microbiol. 1987 Apr;25(4):685–691. doi: 10.1128/jcm.25.4.685-691.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Fennell C. L., Rompalo A. M., Totten P. A., Bruch K. L., Flores B. M., Stamm W. E. Isolation of "Campylobacter hyointestinalis" from a human. J Clin Microbiol. 1986 Jul;24(1):146–148. doi: 10.1128/jcm.24.1.146-148.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fennell C. L., Totten P. A., Quinn T. C., Patton D. L., Holmes K. K., Stamm W. E. Characterization of Campylobacter-like organisms isolated from homosexual men. J Infect Dis. 1984 Jan;149(1):58–66. doi: 10.1093/infdis/149.1.58. [DOI] [PubMed] [Google Scholar]
  9. Fitts R., Diamond M., Hamilton C., Neri M. DNA-DNA hybridization assay for detection of Salmonella spp. in foods. Appl Environ Microbiol. 1983 Nov;46(5):1146–1151. doi: 10.1128/aem.46.5.1146-1151.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gebhart C. J., Edmonds P., Ward G. E., Kurtz H. J., Brenner D. J. "Campylobacter hyointestinalis" sp. nov.: a new species of Campylobacter found in the intestines of pigs and other animals. J Clin Microbiol. 1985 May;21(5):715–720. doi: 10.1128/jcm.21.5.715-720.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gebhart C. J., Ward G. E., Chang K., Kurtz H. J. Campylobacter hyointestinalis (new species) isolated from swine with lesions of proliferative ileitis. Am J Vet Res. 1983 Mar;44(3):361–367. [PubMed] [Google Scholar]
  12. Grimont P. A., Grimont F., Desplaces N., Tchen P. DNA probe specific for Legionella pneumophila. J Clin Microbiol. 1985 Mar;21(3):431–437. doi: 10.1128/jcm.21.3.431-437.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Korolik V., Coloe P. J., Krishnapillai V. A specific DNA probe for the identification of Campylobacter jejuni. J Gen Microbiol. 1988 Feb;134(2):521–529. doi: 10.1099/00221287-134-2-521. [DOI] [PubMed] [Google Scholar]
  15. Lawson G. H., Rowland A. C., Wooding P. The characterisation of Campylobacter sputorum subspecies mucosalis isolated from pigs. Res Vet Sci. 1975 Mar;18(2):121–126. [PubMed] [Google Scholar]
  16. 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]
  17. Ng L. K., Stiles M. E., Taylor D. E. Classification of Campylobacter strains using DNA probes. Mol Cell Probes. 1987 Sep;1(3):233–243. doi: 10.1016/0890-8508(87)90036-3. [DOI] [PubMed] [Google Scholar]
  18. Owen R. J. Nucleic acids in the classification of Campylobacters. Eur J Clin Microbiol. 1983 Aug;2(4):367–377. doi: 10.1007/BF02019473. [DOI] [PubMed] [Google Scholar]
  19. Penner J. L. The genus Campylobacter: a decade of progress. Clin Microbiol Rev. 1988 Apr;1(2):157–172. doi: 10.1128/cmr.1.2.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Picken R. N., Wang Z., Yang H. L. Molecular cloning of a species-specific DNA probe for Campylobacter jejuni. Mol Cell Probes. 1987 Sep;1(3):245–259. doi: 10.1016/0890-8508(87)90037-5. [DOI] [PubMed] [Google Scholar]
  21. Roop R. M., 2nd, Smibert R. M., Johnson J. L., Krieg N. R. DNA homology studies of the catalase-negative campylobacters and "Campylobacter fecalis," an emended description of Campylobacter sputorum, and proposal of the neotype strain of Campylobacter sputorum. Can J Microbiol. 1985 Sep;31(9):823–831. doi: 10.1139/m85-154. [DOI] [PubMed] [Google Scholar]
  22. Roop R. M., 2nd, Smibert R. M., Johnson J. L., Krieg N. R. Differential characteristics of catalase-positive campylobacters correlated with DNA homology groups. Can J Microbiol. 1984 Jul;30(7):938–951. doi: 10.1139/m84-147. [DOI] [PubMed] [Google Scholar]
  23. Rubin F. A., Kopecko D. J., Noon K. F., Baron L. S. Development of a DNA probe to detect Salmonella typhi. J Clin Microbiol. 1985 Oct;22(4):600–605. doi: 10.1128/jcm.22.4.600-605.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  25. Tenover F. C. Diagnostic deoxyribonucleic acid probes for infectious diseases. Clin Microbiol Rev. 1988 Jan;1(1):82–101. doi: 10.1128/cmr.1.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tompkins L. S., Troup N., Labigne-Roussel A., Cohen M. L. Cloned, random chromosomal sequences as probes to identify Salmonella species. J Infect Dis. 1986 Jul;154(1):156–162. doi: 10.1093/infdis/154.1.156. [DOI] [PubMed] [Google Scholar]
  27. Totten P. A., Fennell C. L., Tenover F. C., Wezenberg J. M., Perine P. L., Stamm W. E., Holmes K. K. Campylobacter cinaedi (sp. nov.) and Campylobacter fennelliae (sp. nov.): two new Campylobacter species associated with enteric disease in homosexual men. J Infect Dis. 1985 Jan;151(1):131–139. doi: 10.1093/infdis/151.1.131. [DOI] [PubMed] [Google Scholar]
  28. Totten P. A., Holmes K. K., Handsfield H. H., Knapp J. S., Perine P. L., Falkow S. DNA hybridization technique for the detection of Neisseria gonorrhoeae in men with urethritis. J Infect Dis. 1983 Sep;148(3):462–471. doi: 10.1093/infdis/148.3.462. [DOI] [PubMed] [Google Scholar]

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