文章基本信息

标题：Pyrosequencing with di-base addition for the identification of pathogenic bacteria
本地全文：下载
作者：Dan Pu ; Kunxian Shu
期刊名称：E3S Web of Conferences
印刷版ISSN：2267-1242
电子版ISSN：2267-1242
出版年度：2021
卷号：271
页码：1-6
DOI：10.1051/e3sconf/202127103010
语种：English
出版社：EDP Sciences
摘要：We proposed a novel method for the identification of pathogenic bacteria, in which a partial amplicon of the molecular markers was targeted by using pyrosequencing with di-base addition (PDBA). PDBA was conducted by synchronously adding di-base instead of one base into a reaction, and a set of highly sequence-specific encodings containing the type and the number of incorporated nucleotide(s) (peak height) were obtained. By comparing the encoding sizes of each isolate and the number of incorporated nucleotide(s) in each cycle, moving from first to last, various kinds of bacteria could be unambiguously identified. To verify its feasibility, we simulated PDBA results from thirteen isolates ofMycobacteriumspecies and compared their encoding sizes and the number of incorporated nucleotide(s) in each cycle with those predicted by a homemade software. The thirteen isolates were successfully differentiated. We also targeted partial RNase P RNA gene (rnpB) of culturedM. paratubercuosisandM. celatumto differentiate the two isolates. By comparing the encoding size of each isolate and the number of incorporated nucleotide(s) in each cycle, the twoMycobacteriumisolates were successfully differentiated. In conclusion, PDBA enabled to reliably identify pathogenic bacteria.