Comprehensive and quantitative profiling of microflora of human apical periodontitis

人类根尖周炎微生物菌群的全面定量分析

• 批准号: 17591985
• 负责人: SATO Takuichi
• 金额: $ 1.67万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17591985/
• 关键词: Dentistry; Oral bacteria; Apical periodontitis; PCR; 16S ribosomal RNA

The purpose of the present study was to profile microflora of root canals before and after root canal therapy, using real-time PCR, PCR-cloning, PCR-sequence analyses based on 16S rRNA genes. Informed consent was obtained from each subject and single-rooted teeth with periapical lesions were investigated. Upon access opening, each dentin sample was collected from the root canal. When the periapical lesion healed clinically through chemo-mechanical cleaning and intracanal medication, the root canal dentin sample was obtained again. The quantification of total bacterial DNA was performed by real-time PCR using universal primers based on 16S rRNA genes. PCR products were cloned and partially sequenced. The partial 16S rRNA gene sequences were then compared with those from the GenBank database using the Blast search program through the website of the NCBI (National Center for Biotechnology Information). The concentrations of bacterial DNA after root canal therapy were lower than those upon access opening. The PCR-cloning and PCR-sequence analyses revealed that Fusobacterium were initially predominant, and that Pseudomonas, Bradyrhizobium and Methylobacterium were predominant after root canal therapy. These results indicated that drastic shifts occurred in microflora of root canals by root canal therapy.

本研究采用16SrRNA基因的实时荧光定量PCR、PCR克隆、PCR序列分析等方法，对根管治疗前后根管内的微生物菌群进行分析。获得每个受试者的知情同意书，并对根尖周病变的单根牙进行调查。在进入开口后，从根管中收集每个牙本质样本。当根尖周病变经化学机械清洗和根管内药物治疗临床愈合后，再次获取根管牙本质样本。使用基于16S rRNA基因的通用引物通过实时PCR进行总细菌DNA的定量。对PCR产物进行克隆和部分测序。然后通过NCBI（国家生物技术信息中心）的网站使用Blast搜索程序将部分16S rRNA基因序列与来自GenBank数据库的那些进行比较。根管治疗后的细菌DNA浓度低于根管开放时。PCR克隆和PCR序列分析表明，最初的梭杆菌占主导地位，假单胞菌，慢生根瘤菌和甲基杆菌的根管治疗后占主导地位。结果表明，根管治疗后根管内菌群发生了明显变化。

项目成果

Profiling of dental plaque microflora of root-caries lesions

根龋病变的牙菌斑微生物区系分析

• 发表时间: 2008
• 作者: Fukumoto Y;Yoshioka T;Kobayashi C;Suda H;Kazuhiro Hashimoto
• 通讯作者: Kazuhiro Hashimoto

Detection of periodontopathic bacteria in periodontal pockets by nested polymerase chain reaction

巢式聚合酶链反应检测牙周袋中的牙周病细菌

• 发表时间: 2007
• 作者: Sato T;Abiko Y;Mayanagi G;Matsuyama J and Takahashi N
• 通讯作者: Matsuyama J and Takahashi N

Interface Oral Health Science 2007

• DOI: 10.1007/978-4-431-76690-2
• 发表时间: 2007
• 作者: Makoto Watanabe;O. Okuno;K. Sasaki;N. Takahashi;O. Suzuki;Haruhiko Takada

PCR for detection of mutans streptococci in human dental plaque.

PCR 检测人牙菌斑中的变形链球菌。

• 发表时间: 2005
• 期刊: International Congress Series 1284
• 作者: Miyasawa-Hori H;Miyasawa-Hori H;Hata S;Miyasawa-Hori H;Matsuyama J
• 通讯作者: Matsuyama J

Comparison of age-dependent expression of aggrecan and ADAMTSs in mandibular condylar cartilage, tibial growth plate, and articular cartilage in rats

• DOI: 10.1007/s00418-006-0171-8
• 发表时间: 2006-09-01
• 期刊: HISTOCHEMISTRY AND CELL BIOLOGY
• 影响因子: 2.3
• 作者: Mitani, Hidetoshi;Takahashi, Ichiro;Mitani, Hideo
• 通讯作者: Mitani, Hideo