1993 Fiscal Year Final Research Report Summary
Development and clinical application of detection method of periodontopathic bacteria by using a non-radioactive DNA probe
| Project/Area Number |
03557081
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Conservative dentistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
ISHIKAWA Isao Tokyo Medical and Dental University Faculty of Dentistry Professor, 歯学部, 教授 (10014151)
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| Co-Investigator(Kenkyū-buntansha) |
UMEDA Makoto Tokyo Medical and Dental University Faculty of Dentistry Research Assistant, 歯学部, 助手 (90193937)
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| Project Period (FY) |
1991 – 1993
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| Keywords | periodontal diseases / periodontopathic bacteria / non-radioactive DNA probes / bacterial identification |
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| Research Abstract |
A dot blot DNA hybridization assay for a rapid identification of periodontopathic bacteria in subgingival plaque samples was developed by using bisulfite modified whole genomic DNA probe from which was immunoenzymatically visualized by anti SO_3^- antibody conjugated with alkalinephosphatase. Bisulfite modified DNA probe of P.gingivalis and P.intermedia showed high specificity and sensitivity to detect reference strains. The probe from P.gingivalis was applied on clinical subgingival plaque samples to evaluate the distribution of P.gingivalis in the dentition, and to compare the presence of P.gingivalis with clinical parameters. Fifteen adult periodontitis patients and 6 healthy volunteers were examined. Subgingival plaque samples were taken from 4 sites of all the remaining teeth. At the same time, probing depth and bleeding on probing (BOP) were also recorded. The detection percentage and amounts of P.gingivalis present were statistically compared with probing depth and BOP in the patient group. As a result, P.gingivalis was detected in all patients examined and 3 out of 6 healthy individuals. The detection average was 31% in patient group. When the probing depth was over 4 mm or BOP was positive, the detection percentage of P.gingivalis significantly increased. As more P.gingivalis was identified, the percentage of sites with deep probing depth or that were BOP positive increased significantly. However, P.gingivalis was also detected in clinically healthy sites, and P.gingivalis negative sites with deep probing depth or that were BOP positive were existed in the same patient. These results indicate that P.gingivalis play an important role, but is not the only microorganism responsible for adult periodontitis. Moreover, using RFLPs analysis we examined P.gingivalis strains including clinical isolates on their fimbrillin gene locus. The results indicate that genetic heterogeneity seems to be existing within the fimbrillin gene locus.
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[Publications] Yasui, S., Kojima, T., Hata, S., Zhang, Y.G., Umeda, M.and Ishikawa, I.: "Rapid identification of Porphyromonas gingivalis by bisulfite modified DNA probe method." Journal of Periodontal Research. 28. 98-101 (1993)
Description
「研究成果報告書概要(欧文)」より
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