The development of the species-specific oligonucleotide probes for periodontal pathogens using 16S rRNA sequence

利用 16S rRNA 序列开发牙周病原体物种特异性寡核苷酸探针

• 批准号: 13672155
• 负责人: KEIKO Tsuruda
• 金额: $ 2.24万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672155/
• 关键词: 165 rRNA; Olicionucleotide probe; FISH; Pornhvromonas gingivalis; A. actinomycetemcomitans; 歯周病原性細菌; 16SrRNA; オリゴヌクレオチドプローブ; FISH; In situ hybridization; 検出法

The aim in the development of the species-specific probes is the direct and rapid identification of specific microorganisms in complex subgingival flora. On the basis of 16S rRNA sequences of two Porphyromonas gingivalis strains and three Actinobacillus actinomycetemcomitans strains, two hybridization probes were designed to discriminate between Porphyromonas strains and Prevotella strains and to identify P gingivalis and A. actinomycetemcomitans. For determinative studies, hybridizations with all probes were routinely performed at 45℃. This temperature offered a compromise between specificity and signal intensity. P. gingivalis ATCC33277 and GAI 7802 were strongly fluorescent to the PG177 probe, and A. actinomycetemcomitans Y4 was strongly fluorescent to the AA191 probe. In a mixture of the target strains and F. nucleatum, the use of the species-specific probes detected only the target strains. These results demonstrated the application of this technique to the characterization of complex natural samples.

特异性问题的发展的目的是对复杂的亚gingivalis Flora中特定微生物的直接和快速鉴定。根据两个卟啉单胞菌菌株和三个肌动杆菌的16S rRNA序列，设计了两个杂交问题，以区分卟啉症菌株和prevotella菌株，并识别出斑note虫菌株和识别A. actimanycercetemycetemycetemcetemcetemctemtansans。为了确定研究，常规在45℃以通常进行所有问题的杂交。该温度在特异性和信号强度之间提供了妥协。牙龈疟原虫ATCC33277和GAI 7802强烈荧光到PG177探针，A。放线菌Cetemcomitans Y4强烈荧光到AA191探针。在靶菌株和F.核的混合物中，物种特异性探针的使用仅检测到目标菌株。这些结果证明了该技术在复杂天然样品的表征中的应用。