title: Direct identification of mycobacteria from culture media using a multiplex real-time PCR assay: report on its application in a clinical laboratory in a region of high tuberculosis endemicity.
abstract: There are few commercial assays that easily and correctly identify the mycobacteria from culture in a clinical laboratory with a high workload. Thus, we developed and evaluated a scheme for the identification of mycobacteria using a multiplex real-time PCR assay and report on its application in our laboratory. The scheme consisted of 3 stepwise PCRs. Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) were differentially detected in the step 1 PCR, and the NTM species were identified in the step 2 and 3 PCRs. Over the 1.5-year study period, 1136 isolates of MTC and 618 isolates of NTM were detected, and the species of 608 (98.4%) of the 618 NTM isolates were identified. We conclude that the established scheme is a very useful diagnostic approach for the rapid and accurate identification of MTC and clinically relevant NTM in a clinical laboratory in a region where tuberculosis is endemic.
authors.affiliation: Department of Laboratory Medicine, Gangneung Asan Hospital, University of Ulsan College of Medicine, 415, Bangdong-ri, Sacheon-myeon, Gangeung-si, Gangwon-do, 210-711, Korea.
authors.forename: Choong-Hwan
authors.initials: CH
authors.lastname: Cha
authors.affiliation: Department of Laboratory Medicine, Gangneung Asan Hospital, University of Ulsan College of Medicine, 415, Bangdong-ri, Sacheon-myeon, Gangeung-si, Gangwon-do, 210-711, Korea.
authors.forename: Hae-Kyong
authors.initials: HK
authors.lastname: An
authors.affiliation: Department of Laboratory Medicine, Gangneung Asan Hospital, University of Ulsan College of Medicine, 415, Bangdong-ri, Sacheon-myeon, Gangeung-si, Gangwon-do, 210-711, Korea. Electronic address: jukim@gnah.co.kr.
authors.forename: Jeong-Uk
authors.initials: JU
authors.lastname: Kim
mesh_terms: D019353:Endemic Diseases; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction; D009161:Mycobacterium / Q000235:genetics* / Q000302:isolation & purification*; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D056910:Republic of Korea / Q000453:epidemiology
keywords: Identification of mycobacteria; Multiplex real-time PCR; Mycobacterium tuberculosis complex; Nontuberculous mycobacteria