1887

Abstract

Analysis of the subgingival microflora has recently implicated and several black species in the aetiology of juvenile, adult and rapidly progressing periodontitis. Rapid bacteriological diagnosis has been hampered by the slow growth and fastidious nature of these bacteria. To construct diagnostic probes, dideoxy sequencing of the 16S rRNA molecules from subgroup II, and several closely related species was performed. Next, oligodeoxynucleotides, complementary to defined regions of the 16S rRNA exhibiting considerable evolutionary divergence, were synthesized for use as molecular probes. In a dot-blot hybridization assay, all strains from each of the species for which probes were constructed were correctly identified, with a detection limit of less than 5 10 organisms. No cross-hybridization to closely related species (except for and ) or contaminating bacteria was observed. Using a modified DNA/RNA hybridization technique, the detection could be performed in less than 12 h, as compared to 2-3 weeks using conventional bacteriological procedures.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-7-1931
1988-07-01
2021-10-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/7/mic-134-7-1931.html?itemId=/content/journal/micro/10.1099/00221287-134-7-1931&mimeType=html&fmt=ahah

References

  1. Auffray C., Rougeon F. 1980; Purification of mouse immunological heavy-chain messenger RNAs from total myeloma tumor RNA. European Journal of Biochemistry 107:303–314
    [Google Scholar]
  2. Bonta Y., Zambon J. J., Genco R. J., Neiders M. E. 1985; Rapid identification of periodontal pathogens in subgingival plaque: comparison of indirect immunofluorescence microscopy with bacterial culture for the detection of Actinobacillus actinomycetemcomitans. Journal of Dental Research 64:793–798
    [Google Scholar]
  3. Conner B. J., Reyes A. A., Morin C., Itakura L., Teplitz R. L., Wallace R. B. 1983; Detection of sickle cell #x03B2;s− globulin allele by hybridization with synthetic oligonucleotides. Proceedings of the National Academy of Sciences of the United States of America 80:178–282
    [Google Scholar]
  4. Cristersson L. A., Slots J., Zambon J. J., Genco R. J. 1985; Transmission and colonization of Actinobacillus actinomycetemcomitans in localized juvenile periodontitis. Journal of Periodontology 56:127–131
    [Google Scholar]
  5. Cristersson L. A., Albini B., Zambon J. J., Genco R. J. 1987a; Tissue localization of Actinobacillus actinomycetemcomitans in human periodontitis. I. Light, immunofluorescence, and electron microscope studies. Journal of Periodontology 58:529–539
    [Google Scholar]
  6. Cristersson L. A., WikesjÖ U. M. E., Albini B., Zambon J. J., Genco R. J. 1987b; Tissue localization of Actinobacillus actinomycetemcomitansin human periodontitis. II. Correlation between immunofluorescence and culture techniques. Journal of Periodontology 58:540–545
    [Google Scholar]
  7. Duerden B. I., Goodwin L., O’Neill T. C. A. 1987; Identification of Bacteroides species from adult periodontal disease. Journal of Medical Microbiology 24:133–137
    [Google Scholar]
  8. Eisenstein B. I., Engleberg N. C. 1986; Applied molecular genetics: new tools for microbiologists and clinicians. Journal of Infectious Diseases 153:416–430
    [Google Scholar]
  9. Falkow S. 1985; The use of DNA hybridization for the identification of pathogenic bacteria. In Rapid Methods and Automation in Microbiology and Immunology pp. 30–33 Habermehl K. O. Edited by Berlin: Springer;
    [Google Scholar]
  10. Frederiksen W. 1987; International Committee on Systematic Bacteriology: Subcommittee on Pasteurellaceae and related organisms. International Journal of Systematic Bacteriology 37:474
    [Google Scholar]
  11. French C. K., Savjitt E. D., Simon S. L., Eklund S. M., Chen M. C., Klotz L. C., Vaccaro K. K. 1986; DNA probe detection of periodontal pathogens. Oral Microbiology and Immunology 1:58–62
    [Google Scholar]
  12. GmÜr R., Hrodek K., Saxer U. P., Guggenheim B. 1986; Double-blind analysis of the relation between adult periodontitis and systemic host response to suspected periodontal pathogens. Infection and Immunity 52:768–776
    [Google Scholar]
  13. Göbel U., Gaiser A., Stanbridge E. J. 1987a; Oligonucleotide probes complementary to variable regions of ribosomal RNA discriminate between Mycoplasma species. Journal of General Microbiology 133:1969–1974
    [Google Scholar]
  14. Göbel U., Maas R., Haun G., Vinga-Martins C., Stanbridge E. J. 1987b; Synthetic oligonucleotide probes complementary to ribosomal RNA for group- and species-specific detection of mycoplas- mas. Israel Journal of Medical Sciences 23:742–746
    [Google Scholar]
  15. Göbel U., Maas R., Clad A. 1987c; Quantitative electroelution of oligonucleotides and large DNA fragments from gels, and purification by electrodialysis. Journal of Biochemical and Biophysical Methods 14:245–260
    [Google Scholar]
  16. Gray M. W., Sankoff D., Cedergren R. J. 1984; On the evolutionary descent of organisms and organelles: a global phytogeny based on a highly conserved core structure in small subunit ribosomal RNA. Nucleic Acids Research 12:5837–5852
    [Google Scholar]
  17. Haapasalo M., Ranta H., Ranta K., Shah H. 1986; Black-pigmented Bacteroides spp. in human apical periodontitis. Infection and Immunity 53:149–153
    [Google Scholar]
  18. Haun G., GÖbel U. 1987; Oligonucleotide probes for genus-, species-, and subspecies- specific detection of representatives of the genus Proteus. FEMS Microbiology Letters 43:187–193
    [Google Scholar]
  19. Johnson J. L., Harich B. 1986; Ribosomal ribonucleic acid homology among species of the genus Bacteroides. International Journal of Systematic Bacteriology 36:71–79
    [Google Scholar]
  20. Kuritza A. P., Getty C. E., Shaughnessy P., Hesse R., Salyers A. A. 1986; DNA probes for identification of clinically important Bacteroidesspecies. Journal of Clinical Microbiology 23:343–349
    [Google Scholar]
  21. Lane D. J., Pace B., Olsen G., Stahl D. A., Sogin M. L., Pace N. R. 1985; Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proceedings of the National Academy of Sciences of the United States of America 82:6955–6959
    [Google Scholar]
  22. Lennette E. H., Balows A., Hausler W. J.Jr Shadomy H. J. 1985 Manual of Clinical Microbiology, 7th edn.. Washington, DC:: ASM;
    [Google Scholar]
  23. Miyoshi T., Hanazawa S., Hirose K., Saitoh K., Amano S., Ohmori Y., Kitano S. 1986; Humoral antibody response against Bacteroides gingivalis-specific antigen recognized by monoclonal antibody in adult periodontal patients. Infection and Immunity 53:366–371
    [Google Scholar]
  24. Moore W. E. C. 1987; Microbiology of periodontal disease. Journal of Periodontal Research 22:335–341
    [Google Scholar]
  25. Moore W. E. C., Holdeman L. V., Cato E. P., Good I. J., Smith E. P., Ranney R. R., Palcanis K. G. 1984; Variation in periodontal floras. Infection and Immunity 46:720–726
    [Google Scholar]
  26. Moore W. E. C., Holdeman L. V., Cato E. P., Smibert R. M., Burmeister J. A., Palcanis K. G., Ranney R. R. 1985; Comparative bacteriology of juvenile periodontitis. Infection and Immunity 48:507–519
    [Google Scholar]
  27. Moore L. V. H., Moore W. E. C., Cato E. P., Smibert R. M., Burmeister J. A., Best A. M., Ranney R. R. 1987; Bacteriology of human gingivitis. Journal of Dental Research 66:989–995
    [Google Scholar]
  28. Newman M. G., Socransky S. S. 1987; Predominant cultivable microbiota in periodontitis. Journal of Periodontal Research 12:120–128
    [Google Scholar]
  29. Pace N. R., Stahl D. A., Lane D. J., Olsen G. J. 1986; The analysis of natural microbial populations by ribosomal RNA sequences. In Advances in Microbial Ecology pp. 1–52 Marshall K. C. Edited by New York: Plenum Press;
    [Google Scholar]
  30. Potts T. V., Zambon J. J., Genco R. J. 1985; Reassignment of Actinobacillus actinomycetemcomitans to the genus Haemophilus as Haemophilus actinomycetemcomitans comb.nov. International Journal of Systematic Bacteriology 35:337–341
    [Google Scholar]
  31. Roberts M. C., Moncla B., Kenny G. E. 1987; Chromosomal DNA probes for the identification of Bacteroides species. Journal of General Microbiology 133:1423–1430
    [Google Scholar]
  32. Salyers A. A., Lynn S. P., Gardner J. F. 1983; Use of randomly cloned DNA fragments for identification of Bacteroides thetaiotaomicron. Journal of Bacteriology 154:287–293
    [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74:5463–5467
    [Google Scholar]
  34. Slots J. 1979; Subgingivalmicroflora and periodontal disease. Journal of Clinical Periodontology 6:351–382
    [Google Scholar]
  35. Slots J., Genco R. J. 1984; Black pigmented Bacteroides species, Capnocytophaga species, and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonization, survival, and tissue destruction. Journal of Dental Research 63:412–421
    [Google Scholar]
  36. Slots J., Emrich L. J., Genco R. J., Rosling B. G. 1985; Relationship between some subgingival bacteria and periodontal pocket depth and gain or loss of periodontal attachment after treatment of adult periodontitis. Journal of Clinical Periodontology 12:540–542
    [Google Scholar]
  37. Strzempko M. N., Simon S. L., French C. K., Lippke J. A., Raia F. F., Savitt E. D., Vaccaro K. K. 1987; A cross-reactivity study of whole genomic DNA probes for Haemophilus actinomycetemcomitans, Bacteroides intermedius, and Bacteroides gingivalis. Journal of Dental Research 66:1543–1546
    [Google Scholar]
  38. Viscidi R. P., Yolken R. G. 1987; Molecular diagnosis of infectious diseases by nucleic acid hybridization. Molecular and Cellular Probes 1:3–14
    [Google Scholar]
  39. Weisburg W. G., Oyaizu Y., Oyaizo H., Woese C. R. 1985; Natural relationship between Bacteroides and flavobacteria. Journal of Bacteriology 164:230–236
    [Google Scholar]
  40. Zambon J. J. 1985; Actinobacillus actinomycetemcomitans in human periodontal disease. Journal of Clinical Periodontology 12:1–20
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-7-1931
Loading
/content/journal/micro/10.1099/00221287-134-7-1931
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error