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 Yam, W. C.
Right arrow Articles by Ho, P. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yam, W. C.
Right arrow Articles by Ho, P. L.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, October 2004, p. 4438-4443, Vol. 42, No. 10
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.10.4438-4443.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Direct Detection of Rifampin-Resistant Mycobacterium tuberculosis in Respiratory Specimens by PCR-DNA Sequencing

W. C. Yam,1 C. M. Tam,2 C. C. Leung,2 H. L. Tong,1 K. H. Chan,1 E. T. Y. Leung,1 K. C. Wong,1 W. W. Yew,3 W. H. Seto,1 K. Y. Yuen,1 and P. L. Ho1*

Centre of Infection and Department of Microbiology, Queen Mary Hospital, The University of Hong Kong,1 Tuberculosis and Chest Service, Department of Health,2 Tuberculosis and Chest Unit, Grantham Hospital, Hong Kong Special Administrative Region, People’s Republic of China3

Received 18 April 2004/ Returned for modification 9 June 2004/ Accepted 28 June 2004

This study evaluated the feasibility of a molecular strategy based on identification of Mycobacterium tuberculosis by IS6110 PCR or Cobas Amplicor PCR, and rpoB PCR-DNA sequencing of the 81-bp rifampin resistance determining region (RRDR) for direct detection of rifampin resistance in respiratory specimens. A collection of 2,138 respiratory specimens and 352 nonduplicate M. tuberculosis isolates (including 233 isolates from the evaluated respiratory specimens and an additional collection of 119 stored isolates) from Southern China was investigated. Using culture as the reference test, the overall diagnostic sensitivities of an acid-fast bacillus (AFB) smear, Cobas Amplicor PCR, IS6110 PCR were 54.5% (156 of 286), 86.7% (248 of 286), and 89.2% (255 of 286), respectively. The sensitivities of the rpoB PCR for the specimens with positive AFB smears and with positive PCR results in the IS6110 PCR and/or Cobas Amplicor PCR were 100% (156 of 156) and 92.3% (239 of 259), respectively. Of the 352 nonduplicate M. tuberculosis isolates, the agar proportion method for rifampin reported 39 resistant strains. Full agreement (352 of 352) was found with the agar proportion method and the genotype inferred from the rpoB DNA sequencing data for rifampin. Thirty-nine mutations of nine distinct kinds, eight point mutations, and one deletion within the RRDR were found in the 39 resistant strains. For the direct DNA sequencing performed on rpoB PCR-positive respiratory specimens, the concordance with the agar proportion method and the subsequent PCR-sequencing for the culture isolate was 100%. This strategy has potential application for direct and rapid diagnosis of rifampin-resistant M. tuberculosis in IS6110 PCR or Cobas Amplicor PCR-positive respiratory specimens.

* Corresponding author. Mailing address: Department of Microbiology, University Pathology Building, Queen Mary Hospital, Pokfulam Rd., Hong Kong Special Administrative Region, People’s Republic of China. Phone: 852-28554892. Fax: 852-28551241. E-mail: plho{at}hkucc.hku.hk.

Journal of Clinical Microbiology, October 2004, p. 4438-4443, Vol. 42, No. 10
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.10.4438-4443.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chan, R. C. Y., Hui, M., Chan, E. W. C., Au, T. K., Chin, M. L., Yip, C. K., AuYeang, C. K. W., Yeung, C. Y. L., Kam, K. M., Yip, P. C. W., Cheng, A. F. B. (2007). Genetic and phenotypic characterization of drug-resistant Mycobacterium tuberculosis isolates in Hong Kong. J Antimicrob Chemother 59: 866-873 [Abstract] [Full Text]  
  • Yam, W.-C., Yuen, K.-Y., Kam, S.-Y., Yiu, L.-S., Chan, K.-S., Leung, C.-C., Tam, C.-M., Ho, P.-O., Yew, W.-W., Seto, W.-H., Ho, P.-L. (2006). Diagnostic application of genotypic identification of mycobacteria. J Med Microbiol 55: 529-536 [Abstract] [Full Text]  
  • Leung, E. T. Y., Ho, P. L., Yuen, K. Y., Woo, W. L., Lam, T. H., Kao, R. Y., Seto, W. H., Yam, W. C. (2006). Molecular Characterization of Isoniazid Resistance in Mycobacterium tuberculosis: Identification of a Novel Mutation in inhA. Antimicrob. Agents Chemother. 50: 1075-1078 [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»