Semiquantitative analysis of periodontopathogens by gene amplification

Yoshihisa Sumi, Yoshihisa Yamashita, Yoshio Nakano, Toshihiko Koga

研究成果: ジャーナルへの寄稿記事

抄録

Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis, which are known as major causative organisms of periodontitis, were semiquantitatively identified by two-step polymerase chain reaction (PCR). The sets of specific primers for A. actinomycetemcomitans and P. gingivalis were prepared on the basis of the nucleotide sequences of the IktA and the fimA genes, respectively. The set of universal primers for eubacteria was designed from the nucleotide sequence of a highly conserved region in the eubacterial 16S rRNA sequence. The number of bacteria detectable by one-step PCR assay was no fewer than 103 cells. Less than 103 bacterial cells were detectable by two-step PCR assay. Subgingival plaque samples from 37 sites of 16 patients were obtained with paperpoints and analyzed by two-step PCR assay. More than 2 × 106 bacterial cells were found in the subgingival plaque samples from all diseased sites. In contrast, the number of total bacteria in those from more than half of healthy sites estimated by PCR assay was less than 2 × 106 cells, suggesting that subgingival plaque in diseased sites consists of a relatively larger number of bacteria compared with the population in healthy sites. While A. actinomycetemcomitans was detected in both healthy and diseased sites, P. gingivalis was observed only in diseased sites. Both periodontopathic bacteria occupied a minor part (less than 0.1%) of the total subgingival plaque bacteria.

元の言語英語
ページ(範囲)177-191
ページ数15
ジャーナルJapanese Journal of Medical Science and Biology
48
発行部数4
DOI
出版物ステータス出版済み - 1 1 1995

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Gene Amplification
Polymerase chain reaction
Bacteria
Assays
Aggregatibacter actinomycetemcomitans
Porphyromonas gingivalis
Polymerase Chain Reaction
Nucleotides
Periodontitis
Genes
Population

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

これを引用

Semiquantitative analysis of periodontopathogens by gene amplification. / Sumi, Yoshihisa; Yamashita, Yoshihisa; Nakano, Yoshio; Koga, Toshihiko.

:: Japanese Journal of Medical Science and Biology, 巻 48, 番号 4, 01.01.1995, p. 177-191.

研究成果: ジャーナルへの寄稿記事

Sumi, Yoshihisa ; Yamashita, Yoshihisa ; Nakano, Yoshio ; Koga, Toshihiko. / Semiquantitative analysis of periodontopathogens by gene amplification. :: Japanese Journal of Medical Science and Biology. 1995 ; 巻 48, 番号 4. pp. 177-191.
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abstract = "Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis, which are known as major causative organisms of periodontitis, were semiquantitatively identified by two-step polymerase chain reaction (PCR). The sets of specific primers for A. actinomycetemcomitans and P. gingivalis were prepared on the basis of the nucleotide sequences of the IktA and the fimA genes, respectively. The set of universal primers for eubacteria was designed from the nucleotide sequence of a highly conserved region in the eubacterial 16S rRNA sequence. The number of bacteria detectable by one-step PCR assay was no fewer than 103 cells. Less than 103 bacterial cells were detectable by two-step PCR assay. Subgingival plaque samples from 37 sites of 16 patients were obtained with paperpoints and analyzed by two-step PCR assay. More than 2 × 106 bacterial cells were found in the subgingival plaque samples from all diseased sites. In contrast, the number of total bacteria in those from more than half of healthy sites estimated by PCR assay was less than 2 × 106 cells, suggesting that subgingival plaque in diseased sites consists of a relatively larger number of bacteria compared with the population in healthy sites. While A. actinomycetemcomitans was detected in both healthy and diseased sites, P. gingivalis was observed only in diseased sites. Both periodontopathic bacteria occupied a minor part (less than 0.1{\%}) of the total subgingival plaque bacteria.",
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