Ann Lab Med.  2013 Sep;33(5):349-352. 10.3343/alm.2013.33.5.349.

The Combination of Real-Time PCR and HPLC for the Identification of Non-Tuberculous Mycobacteria

• Park JS
• Choi JI
• Lim JH
• Ahn JJ
• Jegal Y
• Seo KW
• Ra SW
• Jeon JB
• Lee SH
• Kim SR
• Jeong J

• KMID: 1781340
• DOI: http://doi.org/10.3343/alm.2013.33.5.349

Abstract

We used HPLC and AdvanSure real-time PCR (LG Life Sciences, Korea) to retrospectively analyze non-tuberculous mycobacteria (NTM) in 133 clinical specimens. The specimens were culture-positive for NTM and the HPLC method identified 130 strains of mycobacteria from the cultures (97.7%) at the species level. Among the isolates, 48 Mycobacterium. kansasii (36.1%), 39 M. intracellulare (29.3%), 17 M. avium (12.8%), 16 M. abscessus (12.0%), 6 M. fortuitum (4.5%), 2 M. szulgai (1.5%), 2 M. gordonae (1.5%), and 3 unclassified NTM strains (2.3%) were identified. The real-time PCR assay identified 60 NTM-positive specimens (45.1%), 65 negative specimens (48.9%), and 8 M. tuberculosis (TB)-positive specimens (6.0%). The real-time PCR assay is advantageous because of its rapid identification of NTM. However, in our study, the real-time PCR assay showed relatively low sensitivity (45.1%) when using direct specimens including sputum and bronchoalveolar lavage (BAL) fluid. HPLC is useful as it discriminates NTM at the species level, although it is time-consuming and requires specific equipment and technical expertise. A combination of both methods will be helpful for the rapid and accurate identification of mycobacteria in clinical laboratories.

Keyword

Non-tuberculous mycobacteria; Real-time PCR; HPLC

MeSH Terms

Aged
Aged, 80 and over
Bronchoalveolar Lavage Fluid/microbiology
*Chromatography, High Pressure Liquid
DNA, Bacterial/genetics
Female
Humans
Male
Middle Aged
Mycobacterium/*genetics/isolation & purification
Mycobacterium Infections/diagnosis/*microbiology
*Real-Time Polymerase Chain Reaction
Sputum/microbiology
DNA, Bacterial

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