Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probes

被引:132
作者
Kuboniwa, M
Amano, A
Kimura, KR
Sekine, S
Kato, S
Yamamoto, Y
Okahashi, N
Iida, T
Shizukuishi, S
机构
[1] Osaka Univ, Grad Sch Dent, Dept Prevent Dent, Suita, Osaka 5650871, Japan
[2] Osaka Univ, Grad Sch Dent, Dept Oral Frontier Biol, Suita, Osaka 5650871, Japan
[3] Osaka Univ, Dent Hosp, Div Special Care Dent, Suita, Osaka, Japan
[4] Osaka Univ, Microbial Dis Res Inst, Dept Bacterial Infect, Suita, Osaka 565, Japan
来源
ORAL MICROBIOLOGY AND IMMUNOLOGY | 2004年 / 19卷 / 03期
关键词
Actinobacillus actinomycetemcomitans; Porphyromonas gingivalis; Prevotella intermedia; Prevotella nigrescens; quantitative detection; real-time polymerase chain reaction; Tannerella forsythia; TaqMan probe; Treponema denticola;
D O I
10.1111/j.0902-0055.2004.00135.x
中图分类号
R78 [口腔科学];
学科分类号
1003 ;
摘要
Quantitative analysis, with identification of periodontopathic bacteria, is important for the diagnosis, therapeutic evaluation and risk assessment of periodontal disease. We developed a highly sensitive and specific method using real-time polymerase chain reaction (PCR) to detect and quantify six periodontal bacteria: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Treponema denticola, Prevotella intermedia, and Prevotella nigrescens. Species-specific TaqMan probe/primer sets were designed according to 16S ribosomal RNA gene sequences. Plaque and tongue debris specimens were collected from 10 patients with advanced periodontitis and 10 periodontal healthy individuals and analyzed. All species, except for P. nigrescens, were detected in samples from diseased sites in significantly greater numbers than in those from healthy sites, whereas greater numbers of P. nigrescens were found in the controls. These results suggest that the present real-time PCR method with the designed probe/primer sets enabled sensitive detection of the six periodontal bacteria, and may also assist future microbial studies of periodontal diseases.
引用
收藏
页码:168 / 176
页数:9
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