Annals of Clinical Microbiology, The official Journal of the Korean Society of Clinical Microbiology

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Indexed in KCI, KoreaMed, Synapse, DOAJ
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pISSN 2288-0585 eISSN 2288-6850

Mycobacterium tuberculosis and Nontuberculous Mycobacteria by PCR Assay

Original article

Annals of Clinical Microbiology (Ann Clin Microbiol) 2007 October Volume 10, Issue 2, pages 135-142.

https://doi.org/10.5145/ACM.2007.10.2.135

Mycobacterium tuberculosis and Nontuberculous Mycobacteria by PCR Assay

Seong Deok Lee1, Hye Young Lee2, Hyun Chul Kim2, Soo Young Kim1
1Department of Laboratory Medicine, The Catholic University of Korea, St. Vincent’s Hospital, Suwon; 2 Department of Biomedical Life Science, the Graduate School of Health and Environment, Yonsei University, Seoul, Korea

Abstract

Background: The purpose of this study was to evaluate the usefulness of direct PCR for a rapid detection of Mycobacterium tuberculosis (MTB), the differentiation of MTB from nontuberculous mycobacteria (NTM), and the identification of NTM species in acid-fast bacilli (AFB) smear-positive specimens.

Methods: A total of 255 AFB smear-positive respiratory specimens were studied. For the differentiation of MTB from NTM, the bands of the 360 bp, which exists both in MTB and NTM, and the 190 bp, which exists only in MTB, were amplified using the one- tube nested PCR targeting regions of the rpoB gene. The two-tube nested PCR targeting 16S-23S rRNA spacer gene was done with 34 specimens that were negative by one-tube nested PCR. The specimens that were tested positive for NTMs were subsequently subjected to PCR-restriction fragment length polymorphism (PCR-RFLP) analysis based on the rpoB gene for mycobacterial species identification.

Results: Detection rate of MTB and NTM was 87% after the one-tube nested PCR. The detection range of MTB and NTM increased up to 93% after the two-tube nested PCR. The results of PCR-RFLP analysis identified those NTMs as M. avium and M. intracellulare.

Conclusion: This result seems to suggest strongly that the PCR testing especially aiming for differentiation of MTB from NTM, and identificatin of mycobacterial species using AFB smear-positive specimens would be highly importnat in clinical settings for effective treatment of patients. (Korean J Clin Microbiol 2007;10:135-142)

Keywords

MTB, NTM, Acid-Fast Bacilli, PCR, PCR- restriction fragment length polymorphism (PCR-RFLP)