Journal
JOURNAL OF CLINICAL MICROBIOLOGY
Volume 42, Issue 1, Pages 453-457Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JCM.42.1.453-457.2004
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Funding
- NATIONAL INSTITUTE FOR OCCUPATIONAL SAFETY AND HEALTH [R01OH007364] Funding Source: NIH RePORTER
- PHS HHS [1R010H007364-01A1] Funding Source: Medline
- NIOSH CDC HHS [R01 OH007364] Funding Source: Medline
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A single-tube PCR method was developed for efficient identification of nontuberculous mycobacteria (NTM) and their environmental isolates in about 3 111 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 beat 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.
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