title: The combination of real-time PCR and HPLC for the identification of non-tuberculous mycobacteria.
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.
authors.affiliation: Department of Laboratory Medicine, Ulsan University Hospital, University of Ulsan College of Medicine, Ulsan, Korea.
authors.forename: Jae Sun
authors.initials: JS
authors.lastname: Park
authors.forename: Jung-In
authors.initials: JI
authors.lastname: Choi
authors.forename: Ji-Hun
authors.initials: JH
authors.lastname: Lim
authors.forename: Jong-Joon
authors.initials: JJ
authors.lastname: Ahn
authors.forename: Yangjin
authors.initials: Y
authors.lastname: Jegal
authors.forename: Kwang Won
authors.initials: KW
authors.lastname: Seo
authors.forename: Seung Won
authors.initials: SW
authors.lastname: Ra
authors.forename: Jae Bum
authors.initials: JB
authors.lastname: Jeon
authors.forename: Seon Ho
authors.initials: SH
authors.lastname: Lee
authors.forename: Sung-Ryul
authors.initials: SR
authors.lastname: Kim
authors.forename: Joseph
authors.initials: J
authors.lastname: Jeong
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D002851:Chromatography, High Pressure Liquid; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009161:Mycobacterium / Q000235:genetics* / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000175:diagnosis / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction; D013183:Sputum / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial
keywords: HPLC; Non-tuberculous mycobacteria; Real-time PCR