Kim Bum-Joon, Hong Seong-Karp, Lee Keun-Hwa, Yun Yeo-Jun, Kim Eui-Chong, Park Young-Gil, Bai Gil-Han, Kook Yoon-Hoh
Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, Seoul 137-140, Korea.
J Clin Microbiol. 2004 Mar;42(3):1308-12. doi: 10.1128/JCM.42.3.1308-1312.2004.
A novel duplex PCR method that can amplify the 235- and 136-bp rpoB DNAs of Mycobacterium tuberculosis complex and nontuberculous mycobacteria (NTM), respectively, with two different sets of primers was used to differentially identify 44 reference strains and 379 clinical isolates of mycobacteria in a single-step assay. Showing 100% sensitivity and specificity, the duplex PCR method could clearly differentiate M. tuberculosis complex and NTM strains. In addition, restriction fragment length polymorphism analysis and direct sequencing of the amplicon of NTM could be used to supplement species identification.
一种新型双重PCR方法,可分别使用两组不同引物扩增结核分枝杆菌复合群和非结核分枝杆菌(NTM)的235 bp和136 bp rpoB DNA,用于在一步检测中对44株参考菌株和379株分枝杆菌临床分离株进行鉴别。该双重PCR方法具有100%的敏感性和特异性,能够清晰区分结核分枝杆菌复合群和NTM菌株。此外,NTM扩增子的限制性片段长度多态性分析和直接测序可用于补充菌种鉴定。