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Development and Application of Multiprobe Real-Time PCR Method Targeting the hsp65 Gene for Differentiation of Mycobacterium Species from Isolates and Sputum Specimens

Cited 27 time in Web of Science Cited 29 time in Scopus
Authors
Kim, Kijeong; Lee, Hyungki; Lee, Mi-Kyung; Lee, Seoung-Ae; Lim, Seong Yong; Yim, Jae-Joon; Kim, Wonyong; Kook, Yoon-Hoh; Kim, Bum-Joon; Chung, Sang-In; Munkhtsetseg, Bazarragchaa; Koh, Won-Jung; Shim, Tae-Sun
Issue Date
2010-09
Publisher
AMER SOC MICROBIOLOGY
Citation
JOURNAL OF CLINICAL MICROBIOLOGY; Vol.48 9; 3073-3080
Abstract
We developed a multiprobe real-time PCR assay targeting hsp65 (HMPRT-PCR) to detect and identify mycobacterial isolates and isolates directly from sputum specimens. Primers and probes for HMPRT-PCR were designed on the basis of the hsp65 gene sequence, enabling the recognition of seven pathogenic mycobacteria, including Mycobacterium tuberculosis, M. avium, M. intracellulare, M. kansasii, M. abscessus, M. massiliense, and M. fortuitum. This technique was applied to 24 reference and 133 clinical isolates and differentiated between all strains with 100% sensitivity and specificity. Furthermore, this method was applied to sputum specimens from 117 consecutive smear-positive patients with smear results of from a trace to 3+. These results were then compared to those obtained using the rpoB PCR-restriction analysis method with samples from cultures of the same sputum specimens. The HMPRT-PCR method correctly identified the mycobacteria in 89 samples (76.0%, 89/117), and moreover, the sensitivity level was increased to 94.3% (50/53) for sputa with an acid-fast bacillus score equal to or greater than 2+. Our data suggest that this novel HMPRT-PCR method could be a promising approach for detecting pathogenic mycobacterial species from sputum samples and culture isolates routinely in a clinical setting.
ISSN
0095-1137
Language
English
URI
http://hdl.handle.net/10371/77443
DOI
https://doi.org/10.1128/JCM.00939-10
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College of Medicine/School of Medicine (의과대학/대학원)Internal Medicine (내과학전공)Journal Papers (저널논문_내과학전공)
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