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.

本研究的目的是建立一种可用于口腔临床的快速诊断牙周炎的DNA探针方法。该方法最大限度地减少了常规杂交和洗涤的水浴使用，以缩短总时间。样品与煮沸的DNA探针杂交，然后用0.2×SSC/0.1%SDS在75℃下彻底洗涤。检测过程可在2小时内完成。对55例牙周炎患者468个位点的龈下菌斑进行牙周基础治疗前的检测，检测水平均在10个以上。本文对9例牙周炎患者的58个牙周部位进行了洁治和根面平整术后的细菌学和临床观察。将DNA探针法与细菌培养法进行比较，对牙龈卟啉单胞菌的准确率为88%，对牙龈卟啉单胞菌为67%。伴放线菌牙龈卟啉单胞菌的检出率与PD、BOP之间有显著相关性（x2检验，p<0.01）。A. 35岁及以上患者伴放线菌感染与PD的关系（x2检验，P<0.01）。A.放线菌共生菌和BOP的差异不显著。A.伴放线菌在青少年中最高。在青少年牙周浅袋部位（PD<4 mm），未发现牙龈卟啉单胞菌，但有22%的部位存在A.伴放线菌治疗后，仅临床症状消失的部位牙龈卟啉单胞菌的比例显著下降（% 2检验，p<0.01）。然而，A.快速DNA探针法作为一种快速诊断、选择治疗方式和微生物学评价治疗结果的有效工具，具有广阔的应用前景。

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。

Akiyo Komiya: "A rapid DNA robe 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。