Evaluation of periodontal pathogenic microorganisms and periodontitis using rapid DNA probe method.

快速DNA探针法评价牙周病原微生物和牙周炎。

• 批准号: 09671965
• 负责人: TAKAHASHI Junichi
• 金额: $ 1.98万
• 依托单位: TOKYO DENTAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671965/
• 关键词: Rapid DNA probe; Porphyromonas gingivalis; Actinobacillus actinomycelemcomitans; Periodontal disease-associated bacteria; Rapid diagnosis; Periodontal initial therapy; Porphyromonas gingivalis; Actinobacillus actinomycetemcomitans; 歯周炎; 診断法

The aim of the present study was to develop a rapid DNA probe method for the diagnosis of periodontitis that can be used in the dental clinics. The method minimizes the use of water bath for ordinary hybrydization and washing in order to shorten the total time. Samples were hybridized with boiled DNA probe, and then washed thoroughly with 0.2×SSC/0.1% SDS at 75℃. Detection process could be completed within 2 hours. Detection level was more than 10ィイD14ィエD1 bacterial cells/sample.Subgingival plaque samples were taken from 468 sites in 55 patients with periodontitis before the periodontal initial therapy. After the periodontal therapy of scaling and rootplaning, 58 sites in 9 patients with periodontitis were bacteriologically and cliically evaluated. When the DNA probe method was compared with the culture method, the acuracy was 88% for P. gingivalis, 67% for A. actinomycetemcomitans. A significant association was found between the detection of P. gingivalis and PD, BOP (x 2 test, p<0.01). A significant association was also shown between the detection of A. actinomycetemcomitans and PD in patients whose ages were 35 or older (x 2 test, p<0.01). The relationship between A. actinomycetemcomitans and BOP was not significant. The detection rate of A. actinomycetemcomitans was highest in teenagers. At shallow periodontal pocket sites (PD<4mm) in teenagers, no P. gingivalis was found, while 22% of the sites harbored A. actinomycetemcomitans. After the therapy, the proportions of P. gingivalis decreased significantly only in the clinically resolved sites(% 2 test, p<0.01). However, A. actinomycetemcomitans seemed to persist after the therapy.The rapid DNA probe method appears promising as an efficient tool for rapid diagnosis, selection of treatment modalities, and microbiological evaluation of the treatment outcome.

本研究的目的是开发一种可在牙科诊所使用的快速 DNA 探针方法来诊断牙周炎，该方法最大限度地减少了普通杂交和清洗的水浴的使用，以缩短样本的总时间。与煮沸的DNA探针杂交，然后用0.2×SSC/0.1% SDS在75℃下彻底清洗，检测过程可在2小时内完成。 10 D14 D1细菌细胞/样本。在牙周初始治疗前，对55例牙周炎患者的468个部位采集龈下菌斑样本，在牙周洁治和根面平整治疗后，对9例牙周炎患者的58个部位进行细菌学和临床评估。 DNA探针法与培养法对比，对于牙龈卟啉单胞菌准确率为88%， 67% 伴放线菌 A. gingivalis 的检测与 PD、BOP 之间存在显着关联（x 2 检验，p<0.01）。年龄为 35 岁或以上（x 2 检验，p<0.01）。 A. actinomycetemcomitans 与 BOP 之间没有相关性。青少年牙周袋浅部（PD<4mm）检出率最高，但治疗后有22%的部位存在伴放线菌。仅在临床解决部位，牙龈卟啉单胞菌的比例显着下降（% 2 检验，p<0.01）。治疗后伴放线菌似乎仍然存在。快速 DNA 探针方法似乎有望成为快速诊断、选择治疗方式和治疗结果微生物学评估的有效工具。

项目成果

Akiyo Komiya: "A rapid DNA probe method for detection of Porrhyromonas gingivalis and Actinobacillus actinomycetemcomitans"Journal of Periodontology. No.3. (2000)

Akiyo Komiya：“用于检测牙龈卟啉单胞菌和伴放线放线杆菌的快速 DNA 探针方法”牙周病学杂志。

中川種昭,齋藤 淳, 高橋潤一,小宮明代,穂坂康明,山田 了,奥田克爾: "Evalusite Periodontal Testの歯周病関連性細菌の検出能について" 日本歯周病学会誌. 37・2. 312-316 (1995)

Taneaki Nakakawa、Jun Saito、Junichi Takahashi、Akiyo Komiya、Yasuaki Hosaka、Ryo Yamada、Katsuji Okuda：“关于 Evalusite 牙周测试检测牙周病相关细菌的能力”日本牙周病学会杂志 37・2。 312 -316 (1995)

Akiyo Komiya: "A rapid DNA probe method for detection of Porrhyromonas gingivalis and Actinobacillus actinomycetemcomitans"Journal of Periodontology. in press. (2000)

Akiyo Komiya：“用于检测牙龈卟啉单胞菌和伴放线放线杆菌的快速 DNA 探针方法”牙周病学杂志。

Akiyo Komiya: "Development of a Rapid DNA probe method for Porrhyromonas gingivalis and Actinobacillus actinomycetemcomitans and its clinical application"Journal of Japanese Society of Periodontology. 40(2). 162-174 (1998)

Akiyo Komiya：“牙龈卟啉单胞菌和伴放线放线杆菌快速DNA探针方法的开发及其临床应用”日本牙周病学会杂志。

小宮明代: "Porphyromons gingivalisとActinobacillus actinomycetemcomitansに対する簡便DNAプローブ法の開発と臨床応用"日本歯周病学会誌. 40巻2号. 162-174 (1998)

Akiyo Komiya：“牙龈卟啉单胞菌和放线放线杆菌的简单 DNA 探针方法的开发和临床应用”日本牙周病学会杂志，第 40 卷，第 2. 162-174 期（1998 年）。