This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kim, H.-J.
Right arrow Articles by Kim, B.-J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kim, H.-J.
Right arrow Articles by Kim, B.-J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 2006, p. 3855-3862, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.01214-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Differentiation of Mycobacterial Species by hsp65 Duplex PCR Followed by Duplex-PCR-Based Restriction Analysis and Direct Sequencing{triangledown}

Hyun-Ju Kim,1 Ho-Suk Mun,1 Hong Kim,1 Eun-Ju Oh,1 Youngju Ha,2 Gill-Han Bai,3 Young-Gil Park,3 Chang-Yong Cha,1 Yoon-Hoh Kook,1 and Bum-Joon Kim1*

Department of Microbiology and Immunology, Cancer Research Institute and Liver Research Institute, College of Medicine, Seoul National University, Seoul 110-799,1 Mogam Biotechnology Research Institute Diagnostic Lab, Yongin 449-903,2 The Korean Institute of Tuberculosis, The Korean National Tuberculosis Association, Seoul 137-140, South Korea3

Received 13 June 2006/ Returned for modification 6 August 2006/ Accepted 15 August 2006

Here we describe a novel duplex PCR method which can differentiate Mycobacterium tuberculosis and nontuberculosis mycobacteria (NTM) strains by amplifying hsp65 DNAs of different sizes (195 and 515 bp, respectively). The devised technique was applied to 54 reference and 170 clinical isolates and differentiated all strains into their respective groups with 100% sensitivity and specificity. Furthermore, a duplex PCR-restriction analysis (duplex PRA) and a direct sequencing protocol were developed to differentiate NTM strains at the species and subspecies levels based on previously reported hsp65 DNA sequences (H. Kim et al., Int. J. Syst. Evol. Microbiol. 55:1649-1656, 2005) and then applied to 105 NTM clinical isolates. All NTM isolates were clearly differentiated at the species and subspecies levels by subsequent procedures (PRA or direct sequencing) targeting 515-bp NTM duplex PCR amplicons. Our results suggest that novel duplex PCR-based methods are sensitive and specific for identifying mycobacterial culture isolates at the species level.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Cancer Research Institute and Liver Research Institute, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea. Phone: 82 2 740 8316. Fax: 82 2 743 0881. E-mail: kbumjoon{at}snu.ac.kr.

{triangledown} Published ahead of print on 23 August 2006.

Journal of Clinical Microbiology, November 2006, p. 3855-3862, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.01214-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mun, H.-S., Park, J.-H., Kim, H., Yu, H.-K., Park, Y.-G., Cha, C.-Y., Kook, Y.-H., Kim, B.-J. (2008). Mycobacterium senuense sp. nov., a slowly growing, non-chromogenic species closely related to the Mycobacterium terrae complex. Int. J. Syst. Evol. Microbiol. 58: 641-646 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»