The development of the species-specific oligonucleotide probes for periodontal pathogens using 16S rRNA sequence

2003 Fiscal Year Final Research Report Summary

• Project/Area Number: 13672155
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 矯正・小児・社会系歯学
• Research Institution: Hiroshima University
• Principal Investigator: KEIKO Tsuruda Hiroshima University, Graduate school of Biomedical Science, Research Associate, 大学院・医歯薬学総合研究科, 助手 (10112210)
• Project Period (FY): 2001 – 2003
• Keywords: 165 rRNA / Olicionucleotide probe / FISH / Pornhvromonas gingivalis / A. actinomycetemcomitans

Research Abstract

The aim in the development of the species-specific probes is the direct and rapid identification of specific microorganisms in complex subgingival flora. On the basis of 16S rRNA sequences of two Porphyromonas gingivalis strains and three Actinobacillus actinomycetemcomitans strains, two hybridization probes were designed to discriminate between Porphyromonas strains and Prevotella strains and to identify P gingivalis and A. actinomycetemcomitans. For determinative studies, hybridizations with all probes were routinely performed at 45℃. This temperature offered a compromise between specificity and signal intensity. P. gingivalis ATCC33277 and GAI 7802 were strongly fluorescent to the PG177 probe, and A. actinomycetemcomitans Y4 was strongly fluorescent to the AA191 probe. In a mixture of the target strains and F. nucleatum, the use of the species-specific probes detected only the target strains. These results demonstrated the application of this technique to the characterization of complex natural samples.