Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Jan;32(1):75–81. doi: 10.1128/jcm.32.1.75-81.1994

Identification and characterization of genetic cluster groups of Actinobacillus actinomycetemcomitans isolated from the human oral cavity.

J M DiRienzo 1, T L McKay 1
PMCID: PMC262973  PMID: 7907346

Abstract

Actinobacillus actinomycetemcomitans is recognized as a primary pathogen in localized juvenile periodontitis (LJP). Restriction fragment length polymorphisms (RFLP) within a collection of subgingival plaque isolates of this bacterium were identified and characterized as the first step in understanding the pathogenesis of LJP. Over 800 isolates, from members of 18 families (LJP families) with at least one member with active LJP or a documented history of the disease and one or more siblings, less than 13 years of age, having no clinical evidence of LJP and 32 healthy control subjects, were assigned to one of 13 distinct RFLP groups (II to XIV) by using a previously characterized 4.7-kb DNA probe cloned from the reference strain FDC Y4. Isolates belonging to RFLP groups II, IV, V, and XIII predominated subgingival sites in the subjects. Members of RFLP groups II, IV, VII, VIII, X, and XI were recovered only from LJP family subjects, while group XIII and XIV variants were found exclusively in healthy controls. A synthetic oligonucleotide, homologous to the 5' end of the leukotoxin gene (lktA), and the A. actinomycetemcomitans plasmid, pVT745, were tested for their abilities to subdivide the 13 RFLP groups. The leukotoxin probe specifically identified all RFLP group II variants because of the absence of a HindIII site in the upstream noncoding region of the lkt gene complex. The plasmid probe was not as selective but may be useful for identifying clinical isolates belonging to RFLP group I. The use of these probes for the identification of genetic variants of A. actinomycetemcomitans that may be preferentially colonize diseased and healthy subjects will facilitate the study of the role of this important pathogen in periodontal diseases.

Full text

PDF
77

Images in this article

76Image
on p.76
79Image
on p.79
79Image
on p.79

Selected References

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

  1. Anderson D. G., McKay L. L. Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol. 1983 Sep;46(3):549–552. doi: 10.1128/aem.46.3.549-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Asikainen S., Lai C. H., Alaluusua S., Slots J. Distribution of Actinobacillus actinomycetemcomitans serotypes in periodontal health and disease. Oral Microbiol Immunol. 1991 Apr;6(2):115–118. doi: 10.1111/j.1399-302x.1991.tb00462.x. [DOI] [PubMed] [Google Scholar]
  3. Baer P. N. The case for periodontosis as a clinical entity. J Periodontol. 1971 Aug;42(8):516–520. doi: 10.1902/jop.1971.42.8.516. [DOI] [PubMed] [Google Scholar]
  4. Beaty T. H., Boughman J. A., Yang P., Astemborski J. A., Suzuki J. B. Genetic analysis of juvenile periodontitis in families ascertained through an affected proband. Am J Hum Genet. 1987 May;40(5):443–452. [PMC free article] [PubMed] [Google Scholar]
  5. DiRienzo J. M., Cornell S., Kazoroski L., Slots J. Probe-specific DNA fingerprinting applied to the epidemiology of localized juvenile periodontitis. Oral Microbiol Immunol. 1990 Apr;5(2):49–56. doi: 10.1111/j.1399-302x.1990.tb00227.x. [DOI] [PubMed] [Google Scholar]
  6. Gawron-Burke C., Clewell D. B. Regeneration of insertionally inactivated streptococcal DNA fragments after excision of transposon Tn916 in Escherichia coli: strategy for targeting and cloning of genes from gram-positive bacteria. J Bacteriol. 1984 Jul;159(1):214–221. doi: 10.1128/jb.159.1.214-221.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guthmiller J. M., Kolodrubetz D., Cagle M. P., Kraig E. Sequence of the lktB gene from Actinobacillus actinomycetemcomitans. Nucleic Acids Res. 1990 Sep 11;18(17):5291–5291. doi: 10.1093/nar/18.17.5291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Guthmiller J. M., Kolodrubetz D., Kraig E. A panel of probes detects DNA polymorphisms in human and non-human primate isolates of a periodontal pathogen, Actinobacillus actinomycetemcomitans. Microb Pathog. 1993 Feb;14(2):103–115. doi: 10.1006/mpat.1993.1011. [DOI] [PubMed] [Google Scholar]
  9. Guthmiller J. M., Kraig E., Cagle M. P., Kolodrubetz D. Sequence of the lktD gene from Actinobacillus actinomycetemcomitans. Nucleic Acids Res. 1990 Sep 11;18(17):5292–5292. doi: 10.1093/nar/18.17.5292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harshman L., Riley M. Conservation and variation of nucleotide sequences in Escherichia coli strains isolated from nature. J Bacteriol. 1980 Nov;144(2):560–568. doi: 10.1128/jb.144.2.560-568.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kraig E., Dailey T., Kolodrubetz D. Nucleotide sequence of the leukotoxin gene from Actinobacillus actinomycetemcomitans: homology to the alpha-hemolysin/leukotoxin gene family. Infect Immun. 1990 Apr;58(4):920–929. doi: 10.1128/iai.58.4.920-929.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lally E. T., Golub E. E., Kieba I. R., Taichman N. S., Rosenbloom J., Rosenbloom J. C., Gibson C. W., Demuth D. R. Analysis of the Actinobacillus actinomycetemcomitans leukotoxin gene. Delineation of unique features and comparison to homologous toxins. J Biol Chem. 1989 Sep 15;264(26):15451–15456. [PubMed] [Google Scholar]
  13. LeBlanc D. J., Lee L. N., Abu-Al-Jaibat A. R., Sreenivasan P. K., Fives-Taylor P. M. Identification of plasmids in Actinobacillus actinomycetemcomitans and construction of intergeneric shuttle plasmids. Oral Microbiol Immunol. 1993 Apr;8(2):94–99. doi: 10.1111/j.1399-302x.1993.tb00552.x. [DOI] [PubMed] [Google Scholar]
  14. Liljenberg B., Lindhe J. Juvenile periodontitis. Some microbiological, histopathological and clinical characteristics. J Clin Periodontol. 1980 Feb;7(1):48–61. doi: 10.1111/j.1600-051x.1980.tb01948.x. [DOI] [PubMed] [Google Scholar]
  15. Möller A. J. Microbiological examination of root canals and periapical tissues of human teeth. Methodological studies. Odontol Tidskr. 1966 Dec 20;74(5 Suppl):1–380. [PubMed] [Google Scholar]
  16. Newman M. G., Socransky S. S. Predominant cultivable microbiota in periodontosis. J Periodontal Res. 1977 Mar;12(2):120–128. doi: 10.1111/j.1600-0765.1977.tb00114.x. [DOI] [PubMed] [Google Scholar]
  17. Saarela M., Asikainen S., Alaluusua S., Pyhälä L., Lai C. H., Jousimies-Somer H. Frequency and stability of mono- or poly-infection by Actinobacillus actinomycetemcomitans serotypes a, b, c, d or e. Oral Microbiol Immunol. 1992 Oct;7(5):277–279. doi: 10.1111/j.1399-302x.1992.tb00588.x. [DOI] [PubMed] [Google Scholar]
  18. Saxén L. Juvenile periodontitis. J Clin Periodontol. 1980 Feb;7(1):1–19. doi: 10.1111/j.1600-051x.1980.tb01944.x. [DOI] [PubMed] [Google Scholar]
  19. Slots J., Reynolds H. S., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease: a cross-sectional microbiological investigation. Infect Immun. 1980 Sep;29(3):1013–1020. doi: 10.1128/iai.29.3.1013-1020.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Slots J. Selective medium for isolation of Actinobacillus actinomycetemcomitans. J Clin Microbiol. 1982 Apr;15(4):606–609. doi: 10.1128/jcm.15.4.606-609.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Slots J. The predominant cultivable organisms in juvenile periodontitis. Scand J Dent Res. 1976 Jan;84(1):1–10. doi: 10.1111/j.1600-0722.1976.tb00454.x. [DOI] [PubMed] [Google Scholar]
  22. Spitznagel J., Jr, Kraig E., Kolodrubetz D. Regulation of leukotoxin in leukotoxic and nonleukotoxic strains of Actinobacillus actinomycetemcomitans. Infect Immun. 1991 Apr;59(4):1394–1401. doi: 10.1128/iai.59.4.1394-1401.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tanner A. C., Haffer C., Bratthall G. T., Visconti R. A., Socransky S. S. A study of the bacteria associated with advancing periodontitis in man. J Clin Periodontol. 1979 Oct;6(5):278–307. doi: 10.1111/j.1600-051x.1979.tb01931.x. [DOI] [PubMed] [Google Scholar]
  24. Tanner A. C., Visconti R. A., Socransky S. S., Holt S. C. Classification and identification of Actinobacillus actinomycetemcomitans and haemophilus aphrophilus by cluster analysis and deoxyribonucleic acid hybridizations. J Periodontal Res. 1982 Nov;17(6):585–596. doi: 10.1111/j.1600-0765.1982.tb01180.x. [DOI] [PubMed] [Google Scholar]
  25. Wallace R. B., Miyada C. G. Oligonucleotide probes for the screening of recombinant DNA libraries. Methods Enzymol. 1987;152:432–442. doi: 10.1016/0076-6879(87)52050-x. [DOI] [PubMed] [Google Scholar]
  26. Zambon J. J., Christersson L. A., Slots J. Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families. J Periodontol. 1983 Dec;54(12):707–711. doi: 10.1902/jop.1983.54.12.707. [DOI] [PubMed] [Google Scholar]
  27. Zambon J. J., Sunday G. J., Smutko J. S. Molecular genetic analysis of Actinobacillus actinomycetemcomitans epidemiology. J Periodontol. 1990 Feb;61(2):75–80. doi: 10.1902/jop.1990.61.2.75. [DOI] [PubMed] [Google Scholar]

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

RESOURCES