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Development of Pathogenic Bacterial Classification and Identification System based on 16S Ribosomal RNA Sequences

基于16S核糖体RNA序列的病原菌分类鉴定系统开发

基本信息

批准号：
07557028
负责人：
EZAKI Takayuki
金额：
$ 3.84万
依托单位：
Gifu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557028/
关键词：
IDENTIFICATION RIBOSOMAL RNA CLASSIFICATION 16SリボソームRNA

项目摘要

In medical microbiology, identification system of human pathogenic bacteria still depends on conventional phenotypic tests. To overcome this conventional techniques, we aim to accumulate 16S ribosomal RNA sequences of human pathogens and to design their genetic detection and identification system. Including our data, all of the ribosomal RNA sequences of over 200 established class 2 and 3 pathogens were completed until the January of 1997. On the other hand, among 1000 opportunistic human pathogens, 95% of their sequences were determined. Using these data base, we designed rapid genetic detection and identification system ofpathogens within a fewhour. We selectively amplified the position 8 from 5' terminal of 16S rRNA sequence and position 340 to amplify bacterialribosomal DNA sequences. By simply comparing the amplifiedsequenceamong established 1000 pathogens, most human pathogens were able to identify at species level.However, strain differentiationis often requestedin medical microbiology to find sources ofinfection and to determine their pathogenic factors. For this purpose, 16S rRNA data base was not suitable because strain variation werenot observedin the 16S rRNA sequences. IS pattern, SOD gene, and DNA gyrase become candidate for this purpose. Established data set of 16S rRNA sequences are planned to release through internet from our laboratory in very near future.

在医学微生物学中，人类致病菌的鉴定仍依赖于常规的表型试验。为了克服这一传统技术，我们的目标是积累人类病原体的16S核糖体RNA序列，并设计其遗传检测和鉴定系统。包括我们的数据，所有的核糖体RNA序列的200多个已建立的第2类和第3类病原体完成，直到1997年1月。另一方面，在1000种人类机会致病菌中，确定了95%的序列。利用这些数据库，我们设计了一套快速的病原体基因检测和鉴定系统。选择性扩增16SrRNA序列5'端第8位和340位，扩增细菌核糖体DNA序列。通过对1000种已知病原体的序列进行简单比较，大多数人类病原体都能在种水平上进行鉴定，但在医学微生物学中，为了寻找感染源和确定其致病因素，往往需要进行菌株鉴别。16SrRNA数据库不适用于此目的，因为16SrRNA序列中没有包含菌株变异。IS模式、SOD基因和DNA促旋酶成为可能。已建立的16S rRNA序列数据集计划在不久的将来通过互联网从我们的实验室发布。