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RISS 인기검색어
- 검색키워드
- 저자 : Takuichi Sato (검색결과 3 건)
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16S rRNA Genes PCR-RFLP Analysis for Rapid Identification of Oral Anaerobic Gram-Positive Bacilli
Sato, Takuichi,Matsuyama, Junko,Takahashi, Nobuhiro Korean Academy of Oral Biology and the UCLA Dental 2000 International Journal of Oral Biology Vol.25 No.3
Oral anaerobic gram-positive bacilli are frequently isolated from oral infectious sites including carious, endodontic and periodontal lesions. Identification of these bacteria by conventional methods are labor-intensive and time-consuming because of poor growth and non-reactivity in most of the biochemical tests. In this study, in order to develop the rapid identification method for these bacteria, restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal RNA genes〔16S rRNA genes PCR-RFLP〕was applied to anaerobic gram-positive bacilli such as Eubacterium, Mogibacterium and Pseudoramibacter, and the obtained findings were compared with the results of biochemical identification methods. 16S rRNA gene sequences were amplified from isolated genomic DNA samples by the PCR with universal primers. The PCR products were then purified and characterized by single digestion with restriction endonuclease HpaII, and this allowed discrimination among the respective reference strains〔type strains of ATCC〕. In addition, 55 strains were assigned to one of the reference species on the basis of their restriction profiles by digestion with HpaII, and these results were corresponded to those of biochemical identification methods. Furthermore, the 16S rRNA gene sequences were accessed from the GenBank and the RDP data base, and the restriction profiles expected are in accordance with the obtained RFLP patterns in this study. therefore, 16S rRNA genes PCR-RFLP is a rapid and reliable identification method for oral gram-positive bacilli including Eubacterium, Mogibacterium and Pseudoramibacter.
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Matsuyama, Junko,Sato, Takuichi,Takahashi, Nobuhiro Korean Academy of Oral Biology and the UCLA Dental 2000 International Journal of Oral Biology Vol.25 No.3
Restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction〔PCR〕-amplified 16S ribosomal RNA genes〔16S rRNA genes PCR-RFLP〕using MnlI has been shown to be a useful tool in defferentiating oral Actinomyces species. In this study, the method was applied to Actinomyces isolates from human dental plaque〔n=77〕, and the findings were compared with those obtained by biochemical identification methods. As a result, 46 strains out of 55 A. naeslundii genospecies 1 strains were mis-identified as A. israelii〔n=35〕 and A. meyeri〔n=11〕; and 2 strains out of 12 A. naeslundii genospecies 2 strains were mis-identified as A. israelii by biochemical identification methods. Therefore, one should pay attention to the mis-identification of a. naeslundii species by conventional biochemical tests.
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Yasuhiro Ito,Takuichi Sato,Keiko Yamaki,Gen Mayanagi,Kazuhiro Hashimoto,Hidetoshi Shimauchi,Nobuhiro Takahashi 한국미생물학회 2012 The journal of microbiology Vol.50 No.1
This study aimed to profile the microflora in infected root canals before and after root canal treatment using cultureindependent methods. Six infected root canals in singlerooted teeth with periapical lesions from five subjects were included. Quantification of total bacteria was performed by real-time PCR with primers targeting 16S rRNA genes. PCR products with universal 16S rRNA gene primers were cloned and partially sequenced, and bacterial identification at the species level was performed by comparative analysis with the GenBank database. The concentration of extracted DNA before treatment was higher than that after root canal treatment, although the difference was not statistically significant. Sequence analysis revealed that oral bacteria such as Fusobacterium, Streptococcus, Olsenella, and Pseudoramibacter detected in cases before root canal treatment disappeared after treatment. These results suggest that the root canal microflora are distinct before and after root canal treatment, and that treatment changes the microflora in both quantity and quality.
RISS
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