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 Mokrousov, I.
Right arrow Articles by Vyshnevskiy, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mokrousov, I.
Right arrow Articles by Vyshnevskiy, B.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 2002, p. 1617-1620, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1617-1620.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Detection of Ethambutol-Resistant Mycobacterium tuberculosis Strains by Multiplex Allele-Specific PCR Assay Targeting embB306 Mutations

Igor Mokrousov,1* Olga Narvskaya,1 Elena Limeschenko,1 Tatiana Otten,2 and Boris Vyshnevskiy2

Laboratory of Molecular Microbiology, Pasteur Institute, St. Petersburg 197101,1 Laboratory of Microbiology of Tuberculosis, The Research Institute of Phthisiopulmonology, St. Petersburg 193063, Russia2

Received 1 October 2001/ Returned for modification 11 December 2001/ Accepted 12 February 2002

We describe a multiplex allele-specific (MAS)-PCR assay to detect simultaneously mutations in the first and third bases of the embB gene codon 306ATG. These mutations are known to confer ethambutol (EMB) resistance in the majority of clinical Mycobacterium tuberculosis isolates worldwide. The mutated bases are revealed depending on the presence or absence of the respective indicative fragments amplified from the embB306 wild-type allele. Initially optimized on purified DNA samples, the assay was tested on crude cell lysates and auramine-stained sputum slide DNA preparations with the same reproducibility and interpretability of the generated profiles in agarose gel electrophoresis. Since EMB resistance is generally linked to multiple-drug resistance (MDR), the MAS-PCR assay for EMB resistance detection can be used in clinical laboratory practice in areas with a high prevalence and a high transmission rate of MDR-EMB-resistant tuberculosis.

* Corresponding author. Mailing address: Pasteur Institute, 14, Mira St., St. Petersburg, 197101, Russia. Phone: 7-812-233-21-49. Fax: 7-812-232-92-17. E-mail: miv{at}IM4520.spb.edu.

Journal of Clinical Microbiology, May 2002, p. 1617-1620, Vol. 40, No. 5
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.5.1617-1620.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Starks, A. M., Gumusboga, A., Plikaytis, B. B., Shinnick, T. M., Posey, J. E. (2009). Mutations at embB Codon 306 Are an Important Molecular Indicator of Ethambutol Resistance in Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 53: 1061-1066 [Abstract] [Full Text]  
  • Lee, A. S. G., Othman, S. N. K., Ho, Y. M., Wong, S. Y. (2004). Novel Mutations within the embB Gene in Ethambutol-Susceptible Clinical Isolates of Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 48: 4447-4449 [Abstract] [Full Text]  
  • Mokrousov, I., Otten, T., Vyshnevskiy, B., Narvskaya, O. (2003). Allele-Specific rpoB PCR Assays for Detection of Rifampin-Resistant Mycobacterium tuberculosis in Sputum Smears. Antimicrob. Agents Chemother. 47: 2231-2235 [Abstract] [Full Text]  
  • Mokrousov, I., Otten, T., Vyshnevskiy, B., Narvskaya, O. (2002). Detection of embB306 Mutations in Ethambutol-Susceptible Clinical Isolates of Mycobacterium tuberculosis from Northwestern Russia: Implications for Genotypic Resistance Testing. J. Clin. Microbiol. 40: 3810-3813 [Abstract] [Full Text]  
  • Mokrousov, I., Otten, T., Filipenko, M., Vyazovaya, A., Chrapov, E., Limeschenko, E., Steklova, L., Vyshnevskiy, B., Narvskaya, O. (2002). Detection of Isoniazid-Resistant Mycobacterium tuberculosis Strains by a Multiplex Allele-Specific PCR Assay Targeting katG Codon 315 Variation. J. Clin. Microbiol. 40: 2509-2512 [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»