Performance of a real-time PCR assay for the rapid identification of Mycobacterium species

Hye-young Wang; Hyunjung Kim; Sunghyun Kim; Do-kyoon Kim; Sang-Nae Cho; Hyeyoung Lee

doi:10.1007/s12275-015-4495-8

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Research Support, Non-U.S. Gov't
Performance of a real-time PCR assay for the rapid identification of Mycobacterium species: Hye-young Wang ¹, Hyunjung Kim ², Sunghyun Kim ^2,3, Do-kyoon Kim ⁴, Sang-Nae Cho ⁵, Hyeyoung Lee ²; Journal of Microbiology 2015;53(1):38-46.
DOI: https://doi.org/10.1007/s12275-015-4495-8
Published online: January 4, 2015

¹M&D, Inc., Wonju Eco Environmental Technology Center, Wonju 200-722, Republic of Korea, ²Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea, ³Institute for Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea, ⁴Department of Laboratory Medicine, The Catholic University of Korea, St. Vincent’s Hospital, Suwon 442-723, Republic of Korea, ⁵Department of Microbiology, College of Medicine, Yonsei University, Seoul 120-749, Republic of Korea¹M&D, Inc., Wonju Eco Environmental Technology Center, Wonju 200-722, Republic of Korea, ²Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju 220-710, Republic of Korea, ³Institute for Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea, ⁴Department of Laboratory Medicine, The Catholic University of Korea, St. Vincent’s Hospital, Suwon 442-723, Republic of Korea, ⁵Department of Microbiology, College of Medicine, Yonsei University, Seoul 120-749, Republic of Korea

Corresponding author: Hyeyoung Lee , Tel: +82-33-760-2740,

Received: 2 September 2014 • Revised: 27 October 2014 • Accepted: 28 October 2014

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Abstract

Mycobacteria cause a variety of illnesses that differ in severity and public health implications. The differentiation of Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. The diagnosis of diseases caused by NTM is difficult because NTM species are prevalent in the environment and because they have fastidious properties. In the present study, we evaluated 279 clinical isolates grown in liquid culture provided by The Catholic University of Korea, St. Vincent’s Hospital using real-time PCR based on mycobacterial rpoB gene sequences. The positive rate of real-time PCR assay accurately discriminated 100% (195/195) and 100% (84/84) between MTB and NTM species. Comparison of isolates identified using the MolecuTech REBA Myco-ID? and Real Myco-ID? were completely concordant except for two samples. Two cases that were identified as mixed infection (M. intracellulare-M. massiliense and M. avium-M. massiliense co-infection) by PCRREBA assay were only detected using M. abscessus-specific probes by Real Myco-ID?. Among a total of 84 cases, the most frequently identified NTM species were M. intracellulare (n=38, 45.2%), M. avium (n=18, 23.7%), M. massiliense (n=10, 13.2%), M. fortuitum (n=5, 6%), M. abscessus (n=3, 3.9%), M. gordonae (n=3, 3.9%), M. kansasii (n=2, 2.4%), M. mucogenicum (n=2, 2.4%), and M. chelonae (n= 1, 1.2%). Real Myco-ID? is an efficient tool for the rapid detection of NTM species as well as MTB and sensitive and specific and comparable to conventional methods.

Keywords: Mycobacterium tuberculosis (MTB);nontuberculous mycobacteria (NTM);Real Myco-ID?;identification;molecular diagnosis

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Performance of a real-time PCR assay for the rapid identification of Mycobacterium species

J. Microbiol. 2015;53(1):38-46. Published online January 4, 2015

DOI: https://doi.org/10.1007/s12275-015-4495-8

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