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Ribotyping on small-sized spirochetes isolated from subgingival plaque
N E Fiehn1, J M Bangsborg, H Colding
1Department of Oral Microbiology, School of Dentistry, Health Science Faculty, University of Copenhagen, Denmark.
Oral Microbiology and Immunology
|February 1, 1995
Summary
This study used ribotyping to analyze subgingival spirochetes, finding that 1:2:1 spirochetes are more genetically diverse than 2:4:2 spirochetes and Treponema denticola.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Subgingival spirochetes are associated with periodontal diseases.
- Accurate identification and differentiation of spirochete species are crucial for understanding their role in oral health.
Purpose of the Study:
- To differentiate and assess the genetic heterogeneity of subgingival spirochete isolates.
- To compare the genetic diversity of 1:2:1 and 2:4:2 spirochete groups with Treponema socranskii and Treponema denticola.
Main Methods:
- DNA restriction fragment length polymorphism (RFLP) and ribotyping were performed on 17 subgingival spirochete isolates (1:2:1 and 2:4:2), 2 Treponema socranskii strains, and 2 Treponema denticola strains.
- Chromosomal DNA was digested with restriction endonucleases (BamHI, HindIII, PstI, ClaI).
- DNA fragments were separated by agarose gel electrophoresis and hybridized with digoxigenin-labeled 16S and 23S ribosomal RNA probes from Escherichia coli.
Main Results:
- Ribotyping revealed distinct band patterns differentiating the 2:4:2 isolates and T. denticola strains.
- The 1:2:1 spirochete isolates exhibited greater heterogeneity, with up to 11 distinct bands observed after HindIII digestion.
- ClaI digestion separated the 1:2:1 isolates into 8 groups, indicating significant genetic diversity within this group.
Conclusions:
- Ribotyping is an effective method for differentiating subgingival spirochetes.
- The 1:2:1 spirochete group demonstrates higher genetic variability compared to the 2:4:2 group and T. denticola.