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

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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.