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

在医学微生物学中，人类致病菌的鉴定体系仍然依赖于传统的表型试验。为了克服这一传统技术，我们旨在积累人类病原体的16S核糖体RNA序列，并设计其遗传检测和鉴定系统。包括我们的数据在内，到1997年1月，200多种已确定的2类和3类病原体的所有核糖体RNA序列都已完成。另一方面，在1000个机会性人类病原体中，95%的序列被确定。利用这些数据库，我们设计了病原体数小时内的快速基因检测和鉴定系统。我们选择性扩增16S rRNA序列5′端8位和340位，扩增细菌核糖体DNA序列。通过简单地比较已建立的1000种病原体的扩增序列，大多数人类病原体能够在物种水平上识别。然而，在医学微生物学中，经常需要进行菌株鉴别，以发现感染源并确定其致病因素。由于在16S rRNA序列中没有观察到菌株的变异，因此不适合使用16S rRNA数据库。IS模式、SOD基因和DNA旋切酶成为这一目的的候选者。已建立的16S rRNA序列数据集计划在不久的将来从我们的实验室通过互联网发布。

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.