Pyrosequencing with di-base addition for the identification of pathogenic bacteria

We proposed a novel method for the identification of pathogenic bacteria, in which partial amplicon molecular markers was targeted by using pyrosequencing with di-base addition (PDBA). PDBA conducted synchronously adding instead one base into reaction, and set highly sequence-specific encodings containing type number incorporated nucleotide(s) (peak height) were obtained. By comparing encoding sizes each isolate cycle, moving from first to last, various kinds bacteria could be unambiguously identified. To verify its feasibility, we simulated results thirteen isolates Mycobacterium species compared their cycle those predicted homemade software. The successfully differentiated. also RNase P RNA gene ( rnpB ) cultured M. paratubercuosis celatum differentiate two isolates. size In conclusion, enabled reliably identify bacteria.