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Journal of Clinical Microbiology, March 2004, p. 1308-1312, Vol. 42, No. 3
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.3.1308-1312.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Differential Identification of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria by Duplex PCR Assay Using the RNA Polymerase Gene (rpoB)

Bum-Joon Kim,1 Seong-Karp Hong,{dagger} Keun-Hwa Lee,1 Yeo-Jun Yun,1 Eui-Chong Kim,2 Young-Gil Park,3 Gil-Han Bai,3 and Yoon-Hoh Kook1*

Departmentof Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, and Clinical Research Institute,1 Department of Clinical Pathology, Seoul National University College of Medicine, Seoul 110-799,2 The Korean Institute of Tuberculosis, Korean National Tuberculosis Association, Seoul 137-140,Korea3

Received 8 July 2003/ Returned for modification 20 August 2003/ Accepted 18 November 2003

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.

* Corresponding author. Mailing address: Department of Microbiology, Seoul National University College of Medicine, 28 Yongon-Dong, Chongno-Gu, Seoul 110-799, Korea. Phone: 82-2-740-8306. Fax: 82-2-743-0881. E-mail: yhkook{at}plaza.snu.ac.kr.

{dagger} Present address: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea.

Journal of Clinical Microbiology, March 2004, p. 1308-1312, Vol. 42, No. 3
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.3.1308-1312.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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