Korean J Lab Med.  2008 Apr;28(2):103-108. 10.3343/kjlm.2008.28.2.103.

Detection of Mycobacterium tuberculosis complex Using Real-time Polymerase Chain Reaction

• Chang HE
• Heo SR
• Yoo KC
• Song SH
• Kim SH
• Kim HB
• Park KU
• Song J
• Lee JH
• Park SS
• Kim EC

• KMID: 854836
• DOI: http://doi.org/10.3343/kjlm.2008.28.2.103

Abstract

BACKGROUND: For the detection of Mycobacterium tuberculosis complex (MTB), PCR is known to be sensitive, specific, and rapid compared to the conventional methods of acid-fast-bacilli (AFB) smear and culture. We evaluated a new approach for MTB detection using real-time PCR. METHODS: The specificity of real-time PCR was evaluated using 20 MTB isolates and 37 nontuberculous mycobacteria (NTM) isolates identified by AccuProbe Mycobacterium tuberculosis complex colony identification test (Gen-Probe Inc., USA) and Myco-ID (M&D, Korea). One hundred sputum specimens (50 AFB smear-positive and 50 negative specimens) were analyzed using real-time PCR and Amplicor Mycobacterium tuberculosis test (Roche, Germany). The results of real-time PCR positives (55 samples) and negatives (598 samples) were analyzed by AFB smear and culture. RESULTS: The real-time PCR assay accurately discriminated between MTB and NTM species. Realtime PCR and Amplicor test yielded the same results in 96.0% (96/100) of the sputum specimens tested. The sensitivity and specificity of real-time PCR based on AFB culture were 97.4% and 88.5%, respectively. Of the 55 real-time PCR positive specimens, 83.6% (46/55) were culture-positive, 30.9% (17/55) were smear-positive, 52.7% (29/55) were smear-negative and culture-positive, and 14.5% (8/55) were both smear and culture-negative. Among the 598 real-time PCR negative specimens, 60 were not tested for AFB smear or culture and 10 were contaminated. Of the remaining 528 specimens, 478 (90.5%) were both smear and culture-negative and 39 (7.4%) were culture-positive. CONCLUSIONS: For the detection of MTB, real-time PCR was sensitive and specific and comparable to conventional methods. It can be used for rapid identification of M. tuberculosis in clinical laboratories.

Keyword

Mycobacterium tuberculosis complex; Real-time polymerase chain reaction; Sensitivity; Specificity

MeSH Terms

*Bacterial Typing Techniques
Computer Systems
Humans
Mycobacterium tuberculosis/classification/genetics/*isolation & purification
Polymerase Chain Reaction/*methods
Reagent Kits, Diagnostic
Sensitivity and Specificity
Tuberculosis/*microbiology

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