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

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, January 2004, p. 453-457, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.453-457.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a Single-Tube, Cell Lysis-Based, Genus-Specific PCR Method for Rapid Identification of Mycobacteria: Optimization of Cell Lysis, PCR Primers and Conditions, and Restriction Pattern Analysis

Molecular Toxicology Division, Department of Environmental Health, University of Cincinnati Medical Center, Cincinnati, Ohio 45267-0056,

Received 5 May 2003/ Returned for modification 4 June 2003/ Accepted 29 September 2003

A single-tube PCR method was developed for efficient identification of nontuberculous mycobacteria (NTM) and their environmental isolates in about 3 h without conventional DNA isolation. The following three steps were optimized or developed: (i) a simple, 6-min direct cell lysis protocol as a PCR prestep for generation of DNA-template, (ii) an improved Mycobacterium-specific PCR amplification protocol with a broader species specificity using newly designed primers targeting a 228-bp region of the 65-kDa heat shock protein (hsp) gene and optimal PCR amplification conditions, and (iii) a genus-specific restriction analysis of the PCR product for conclusive identification of the unknown NTM isolates.

This article has been cited by other articles:

• Elbir, H., Abdel-Muhsin, A.-M., Babiker, A. (2008). A One-Step DNA PCR-based Method for the Detection of Mycobacterium tuberculosis Complex Grown on Lowenstein-Jensen Media. Am J Trop Med Hyg 78: 316-317 [Abstract] [Full Text]  
• Richter, E., Rusch-Gerdes, S., Hillemann, D. (2006). Evaluation of the GenoType Mycobacterium Assay for Identification of Mycobacterial Species from Cultures.. J. Clin. Microbiol. 44: 1769-1775 [Abstract] [Full Text]  
• Khan, I. U. H., Selvaraju, S. B., Yadav, J. S. (2005). Method for Rapid Identification and Differentiation of the Species of the Mycobacterium chelonae Complex Based on 16S-23S rRNA Gene Internal Transcribed Spacer PCR-Restriction Analysis. J. Clin. Microbiol. 43: 4466-4472 [Abstract] [Full Text]  
• Aldous, W. K., Pounder, J. I., Cloud, J. L., Woods, G. L. (2005). Comparison of Six Methods of Extracting Mycobacterium tuberculosis DNA from Processed Sputum for Testing by Quantitative Real-Time PCR. J. Clin. Microbiol. 43: 2471-2473 [Abstract] [Full Text]