1996 Fiscal Year Final Research Report Summary
Development of non-radioactive DNA probe and its clinical use
| Project/Area Number |
06557101
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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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 |
WATANABE Hisashi Tokyo Medical and Dental University, Dental Asso.Prof, 歯学部, 助教授 (40143606)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIYAMA Eiichi Tokyo Medical and Dental University, Dental Research Asso., 歯学部, 助手 (20242208)
HAGIWARA Satsuki Tokyo Medical and Dental University, Dental Lecturer, 歯学部, 講師 (70134715)
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| Project Period (FY) |
1994 – 1996
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| Keywords | AP-PCR / genotype / primer / Periodontopathic bacteria / non-radioactive DNA probe |
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| Research Abstract |
Actinobacillus actinomycetemcomitans (A.a..) is one of the important pathogens in the periodontal diseases. Recently, Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) has been proposed for detecting genetic polymorphism without prior knowledge of the DNA sequence. In this study, we examined the utility of AP-PCR for genotyping of A.a.. Whole genomic DNA from 8 laboratory strains was extracted by a modification of the method of Van Steenberen. A single 10-mer, synthetic, oligonucleotide of random sequence was used as primer in AP-PCR.One (GAAACGGGTG) of the 20 primers (Operon 10-mer kit A) was found useful in differentiating these strains. The result indicates that AP-PCR is capable of distinguishing different strains of A.a... In addition, we evaluated the potential of application of non-radioactive DNA probe method (Affirm DP^<TM>) in clinical diagnosis and evaluation of treatment efficacy. Sensitivity and specificity of cut-off level (10^4) of the method with bacterial culture in detection of Bacteroide forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were compared. The result showed that detection percentage of DNA probe method with 10^4 cut-off was equal to or higher than culture method for B.forsythus and P.gingivalis. It is suggested to be a useful chair-side DNA probe kit as an aid in clinical diagnosis and evaluation of treatment efficacy.
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