Establishment of rapid method for detection of periodontitis-associated microorganisms

牙周炎相关微生物快速检测方法的建立

• 批准号: 06557102
• 负责人: MURAYAMA Yoji
• 金额: $ 5.63万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557102/
• 关键词: Periodontitis-associated microorganisms; 16S rRNA; PcR; 歯科臨床; 細菌検査システム; 16S rRNA; 歯周病関連細菌; 歯周病; 歯周病原性細菌; リボゾームRNA; 細菌検査

The goal of this project is to establish the microbiological test that can be used for dental treatments, especially for the periodontal treatments by general dentists. This project consists of the following three parts : First study indicated the complexity of putative pathogens relating to periodontitis. In the latter two studies, we established the practical methods for rapid and sensitive detection of multiple pathogens and carried out the test for patients visiting general medical facilities.1) Analyzing the relationships of specific microorganisms in periodontitis and its related diseases.To analyze the microbiological aspects for etiology of periodontitis, gingival hyperemia and endodontic-periodontic lesions, we detected specific microorganisms in subgingival pockets or in root canals. We selected family cases that each member manifested an early-onset periodontitis because of their genetic homogeneity for host defense functions, and acquisition and transmission of pathogens. T … More he results indicated that the possible roles of specific pathogens in each case. Their distributions, however, did not correlate with clinical diagnosis, and even the same family member had differnt pathogens. In conclusion detection of multiple pathogens is needed for assessing the treatment of periodontitis.2) Establishment of rapid method for detection of pathogens.We established the practical methods for detection of the multiple pathogens by easy and rapid procedures. The principle for the detection of microorganisms was based on differences of the sequence in bacterial 16S ribosomal RNA (rRNA). Direct detection of 16S rRNA by hybridization had low sensitivity and also decreased sensitivity during keeping the samples. Therefore we developed two kind of methods, the colony-hybridization method that combine with culture method and the 16S rRNA-based polymerase chain reaction (PCR) method that amplify the 16S rRNA gene. Both methods showed satisfying sensitivity and accuracy for detection of specific pathogens from crude plaque samples. Especially the 16S rRNA-based PCR was rapid and was easy to perform.3) Application of the microbiological test in general medical facilities.We analyze the microflora of subgingival plaques from the patients of insulin-dependent diabetes mellitus and leprosy that visited the clinics in medical school or sanatorium. Less

本项目的目标是建立可用于牙科治疗，特别是普通牙科医生牙周治疗的微生物检测。本课题主要包括以下三个部分：第一部分研究表明牙周炎致病菌的复杂性。在后两项研究中，我们建立了实用的快速、敏感的多病原体检测方法，并对普通医疗机构就诊的患者进行了试验。1）分析牙周炎及其相关疾病中特定微生物的关系。分析牙周炎、牙龈充血和牙髓牙周病变的病原学微生物学方面，我们在龈下袋或根管内检测到特定的微生物。我们选择的家庭病例，每个成员表现出早发性牙周炎，因为他们的宿主防御功能的遗传同质性，病原体的获取和传播。不 ...更多信息 结果表明，在每种情况下，特定病原体的可能作用。但它们的分布与临床诊断无关，甚至同一家族成员有不同的病原体。结论牙周炎治疗效果的评价需要多种病原体的检测。2）建立牙周炎病原体快速检测方法。微生物检测的原理是基于细菌16S核糖体RNA（rRNA）序列的差异。16SrRNA直接杂交检测的灵敏度较低，而且在保存样品期间灵敏度也会降低。为此，我们发展了两种方法，即联合收割机与培养法相结合的菌落杂交法和基于16SrRNA的PCR扩增法。这两种方法显示出令人满意的敏感性和准确性检测特定病原体从天然菌斑样品。特别是16SrRNA PCR法具有快速、简便的特点。3）微生物检测在一般医疗机构中的应用对医学院门诊或疗养院就诊的胰岛素依赖型糖尿病和麻风病人的龈下菌斑进行了菌群分析。少

项目成果

Isoshima,O: "Distribution of black-pigmented prevotella and Porphyromonas species in the dentition of moderate periodon titis patients." Microbial Ecology in Healt and Disease. 8. 159-169 (1995)

Isoshima,O：“中度牙周炎患者牙列中黑色素普雷沃氏菌和卟啉单胞菌的分布。”

Kurihara,H.: "Amicrobiological and mmunological study of endodontic-perioclontic lesions" Journal of Endodontics. 21. 617-621 (1995)

Kurihara,H.：“牙髓-阵周病变的微生物学和免疫学研究”牙髓学杂志。

Ohyama, H., Takashiba, S., Oyaizu, K., Nagai, A., Naruse, T., Inoko, H., Kurihara, H.and Murayama, Y.: "HLA class II genotypes associated with early-onset periodontitis : DQB1 molecule primarily confers susuceptibility to the disease." Journal of Periodon

Ohyama, H.、Takashiba, S.、Oyaizu, K.、Nagai, A.、Naruse, T.、Inoko, H.、Kurihara, H. 和 Murayama, Y.：“与早发型相关的 HLA II 类基因型

村山洋二: "治療から予防への始動-序にかえて-,特集/見えてきた遺伝子診断の全貌" 歯界展望. 84. 818-823 (1994)

Yoji Murayama：“从治疗到预防——而不是开始——，特辑/揭示基因诊断的全貌”Dental Perspective 84. 818-823 (1994)。

Takahashi,K: "Clinical and laboratory studies on a patient with early.on set periodontitis and her family members" Journal of Periodontology. 66. 403-412 (1995)

Takahashi,K：“早期牙周炎患者及其家人的临床和实验室研究”牙周病学杂志。