Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Feb;27(2):321–326. doi: 10.1128/jcm.27.2.321-326.1989

Identification and classification of Campylobacter strains by using nonradioactive DNA probes.

D Chevrier 1, D Larzul 1, F Megraud 1, J L Guesdon 1
PMCID: PMC267300  PMID: 2915027

Abstract

Acetylaminofluorene-labeled genomic DNA probes were used for the identification and classification of Campylobacter strains. Relationships among 17 well-known strains of Campylobacter species and subspecies were studied by comparing acetylaminofluorene- or 32P-labeled probes. Results obtained with both methods were closely correlated and were in agreement with those already reported. In an identification experiment, hybridization with nonradioactive probes was performed on 60 strains isolated from stool samples after subculturing and quick DNA extraction; conventional biochemical tests were conducted in parallel. A good correlation was found between the results obtained by nonradioactive hybridization and by biochemical tests. Thus, the acetylaminofluorene-labeled genomic DNA probe method is an interesting alternative for laboratories without access to radioisotopes for the identification and classification of bacteria.

Full text

PDF
321

Images in this article

Selected References

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

  1. Altwegg M., Burnens A., Zollinger-Iten J., Penner J. L. Problems in identification of Campylobacter jejuni associated with acquisition of resistance to nalidixic acid. J Clin Microbiol. 1987 Sep;25(9):1807–1808. doi: 10.1128/jcm.25.9.1807-1808.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  3. Dutilh B., Bébéar C., Taylor-Robinson D., Grimont P. A. Detection of Chlamydia trachomatis by in situ hybridization with sulphonated total DNA. Ann Inst Pasteur Microbiol. 1988 Jan-Feb;139(1):115–127. doi: 10.1016/0769-2609(88)90099-3. [DOI] [PubMed] [Google Scholar]
  4. Edelstein P. H. Evaluation of the Gen-Probe DNA probe for the detection of legionellae in culture. J Clin Microbiol. 1986 Mar;23(3):481–484. doi: 10.1128/jcm.23.3.481-484.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Fuchs R., Daune M. Physical basis of chemical carcinogenesis by N-2-fluorenylacetamide derivatives and analogs. FEBS Lett. 1973 Aug 15;34(2):295–298. doi: 10.1016/0014-5793(73)80815-4. [DOI] [PubMed] [Google Scholar]
  7. Fuchs R., Daune M. Physical studies on deoxyribonucleic acid after covalent binding of a carcinogen. Biochemistry. 1972 Jul 4;11(14):2659–2666. doi: 10.1021/bi00764a017. [DOI] [PubMed] [Google Scholar]
  8. Hyman H. C., Yogev D., Razin S. DNA probes for detection and identification of Mycoplasma pneumoniae and Mycoplasma genitalium. J Clin Microbiol. 1987 Apr;25(4):726–728. doi: 10.1128/jcm.25.4.726-728.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hébert G. A., Edmonds P., Brenner D. J. DNA relatedness among strains of Campylobacter jejuni and Campylobacter coli with divergent serogroup and hippurate reactions. J Clin Microbiol. 1984 Jul;20(1):138–140. doi: 10.1128/jcm.20.1.138-140.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kiehn T. E., Edwards F. F. Rapid identification using a specific DNA probe of Mycobacterium avium complex from patients with acquired immunodeficiency syndrome. J Clin Microbiol. 1987 Aug;25(8):1551–1552. doi: 10.1128/jcm.25.8.1551-1552.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Landegent J. E., Jasen in de Wal N., Baan R. A., Hoeijmakers J. H., Van der Ploeg M. 2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences. Exp Cell Res. 1984 Jul;153(1):61–72. doi: 10.1016/0014-4827(84)90448-8. [DOI] [PubMed] [Google Scholar]
  12. Larzul D., Thiers V., Courouce A. M., Bréchot C., Guesdon J. L. Non-radioactive hepatitis B virus DNA probe for detection of HBV-DNA in serum. J Hepatol. 1987 Oct;5(2):199–204. doi: 10.1016/s0168-8278(87)80573-1. [DOI] [PubMed] [Google Scholar]
  13. Le Guern A., Leng M., Kourilsky P. Monoclonal antibodies to DNA modified by the carcinogen N-acetoxy-N-2 acetyl aminofluorene. Dev Biol Stand. 1984;57:409–414. [PubMed] [Google Scholar]
  14. Masse M. J., Meulien P., Le Guern A., Kourilsky P. Monoclonal antibody detection of 2-acetyl-aminofluorene-modified DNA probes for the specific detection of nucleic acids in hybridization procedures. Ann Inst Pasteur Immunol. 1985 Nov-Dec;136D(3):231–243. doi: 10.1016/s0769-2625(85)80109-4. [DOI] [PubMed] [Google Scholar]
  15. Megraud F., Bonnet F., Garnier M., Lamouliatte H. Characterization of "Campylobacter pyloridis" by culture, enzymatic profile, and protein content. J Clin Microbiol. 1985 Dec;22(6):1007–1010. doi: 10.1128/jcm.22.6.1007-1010.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Megraud F., Bonnet F. Unusual campylobacters in human faeces. J Infect. 1986 May;12(3):275–276. doi: 10.1016/s0163-4453(86)94398-7. [DOI] [PubMed] [Google Scholar]
  17. Miller E. C., Juhl U., Miller J. A. Nucleic acid guanine: reaction with the carcinogen N-acetoxy-2-acetylaminofluorene. Science. 1966 Sep 2;153(3740):1125–1127. doi: 10.1126/science.153.3740.1125. [DOI] [PubMed] [Google Scholar]
  18. Moore R. L. Nucleic acid reassociation as a guide to genetic relatedness among bacteria. Curr Top Microbiol Immunol. 1974;64(0):105–128. doi: 10.1007/978-3-642-65848-8_4. [DOI] [PubMed] [Google Scholar]
  19. Moseley S. L., Huq I., Alim A. R., So M., Samadpour-Motalebi M., Falkow S. Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis. 1980 Dec;142(6):892–898. doi: 10.1093/infdis/142.6.892. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. 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]
  24. Roberts M. C., Hillier S. L., Schoenknecht F. D., Holmes K. K. Nitrocellulose filter blots for species identification of Mobiluncus curtisii and Mobiluncus mulieris. J Clin Microbiol. 1984 Oct;20(4):826–827. doi: 10.1128/jcm.20.4.826-827.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Roberts M. C., Moncla B., Kenny G. E. Chromosomal DNA probes for the identification of Bacteroides species. J Gen Microbiol. 1987 Jun;133(6):1423–1430. doi: 10.1099/00221287-133-6-1423. [DOI] [PubMed] [Google Scholar]
  26. Romaniuk P. J., Zoltowska B., Trust T. J., Lane D. J., Olsen G. J., Pace N. R., Stahl D. A. Campylobacter pylori, the spiral bacterium associated with human gastritis, is not a true Campylobacter sp. J Bacteriol. 1987 May;169(5):2137–2141. doi: 10.1128/jb.169.5.2137-2141.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. 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]
  30. Steele T. W., Sangster N., Lanser J. A. DNA relatedness and biochemical features of Campylobacter spp. isolated in central and South Australia. J Clin Microbiol. 1985 Jul;22(1):71–74. doi: 10.1128/jcm.22.1.71-74.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tchen P., Fuchs R. P., Sage E., Leng M. Chemically modified nucleic acids as immunodetectable probes in hybridization experiments. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3466–3470. doi: 10.1073/pnas.81.11.3466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Terpstra W. J., Schoone G. J., Ligthart G. S., ter Schegget J. Detection of Leptospira interrogans in clinical specimens by in situ hybridization using biotin-labelled DNA probes. J Gen Microbiol. 1987 Apr;133(4):911–914. doi: 10.1099/00221287-133-4-911. [DOI] [PubMed] [Google Scholar]
  33. Tompkins L. S., Krajden M. Approaches to the detection of enteric pathogens, including Campylobacter, using nucleic acid hybridization. Diagn Microbiol Infect Dis. 1986 Mar;4(3 Suppl):71S–78S. doi: 10.1016/s0732-8893(86)80044-x. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Totten P. A., Patton C. M., Tenover F. C., Barrett T. J., Stamm W. E., Steigerwalt A. G., Lin J. Y., Holmes K. K., Brenner D. J. Prevalence and characterization of hippurate-negative Campylobacter jejuni in King County, Washington. J Clin Microbiol. 1987 Sep;25(9):1747–1752. doi: 10.1128/jcm.25.9.1747-1752.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES