The Combination of Real-Time PCR and HPLC for the Identification of Non-Tuberculous Mycobacteria
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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.
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.
참고문헌 (16)
- "Management of opportunist mycobacterial infections: Joint Tuberculosis Committee Guidelines 1999. Subcommittee of the Joint Tuberculosis Committee of the British Thoracic Society", Thorax, 55, pp.210~218 (2000)
- Daley CL, "Nontuberculous mycobacterial disease in transplant recipients: early diagnosis and treatment", Curr Opin Organ Transplant, 14, pp.619~624 (2009)
- Corbett EL, "The growing burden of tuberculosis: global trends and interactions with the HIV epidemic", Arch Intern Med, 163, pp.1009~1021 (2003)
- Falkinham JO 3rd, "Nontuberculous mycobacteria in the environment", Clin Chest Med, 23, pp.529~551 (2002)
- 왕혜영, "도말 양성 임상검체에서 MolecuTech Real MTB-ID을 이용한 결핵균과 비결핵균의 검출평가", Annals of Clinical Microbiology, 14(3), pp.103~109 (2011), 대한임상미생물학회
- 황상선, "LG AdvanSure TB", NTM Real-Time PCR Kit의 항산균 진단 수행 능력 평가, 2011, pp.14(2), 55~59[DOI]
- 박철민, "중합효소연쇄반응-제한절편길이다형성을 이용한 비결핵항산균의 분리", Annals of Laboratory Medicine, 26(3), pp.161~167 (2006), 대한진단검사의학회
- Burman WJ, "Review of false-positive cultures for Mycobacterium tuberculosis and recommendations for avoiding unnecessary treatment", Clin Infect Dis, 31, pp.1390~1395 (2000)
- 정윤성, "5년간 울산대학교병원에서 HPLC법을 이용한 Mycobacteria의 동정과 균종 분포", Annals of Laboratory Medicine, 28(1), pp.24~33 (2008), 대한진단검사의학회
- Butler WR, "Mycolic acid analysis by high-performance liquid chromatography for identification of Mycobacterium species", Clin Microbiol Rev, 14, pp.704~726 (2001)
- Jeong J, "The Use of High Performance Liquid Chromatography to Speciate and Characterize the Epidemiology of Mycobacteria", LABMEDICINE, 42(10), pp.612~617 (2011), AMER SOC CLINICAL PATHOLOGY
- Clinical and Laboratory Standards Institute, "Laboratory detection and identification of mycobacteria; approved guideline M48-A" (2008), CLSI
- Shinnick TM, "National plan for reliable tuberculosis laboratory services using a systems approach. Recommendations from CDC and the Association of Public Health Laboratories Task Force on Tuberculosis Laboratory Services", MMWR Recomm Rep, 54, pp.1~12 (2005)
- 장호은, "실시간-중합효소연쇄반응을 이용한 결핵균의 검출", Annals of Laboratory Medicine, 28(2), pp.103~108 (2008), 대한진단검사의학회
- 김영진, "호흡기 검체를 이용한 AdvanSure TB", NTM Real Time PCR Kit의 항산균 진단 능력평가, 2008, pp.28(1), 34~38[DOI]
- Koh WJ, "Clinical significance of nontuberculous mycobacteria isolated from respiratory specimens in Korea", Chest, 129, pp.341~348 (2006)