Species spectrum of nontuberculous mycobacteria isolated from suspected tuberculosis patients,identification by multi locus sequence analysis |
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| Institution: | 1. Department of Clinical Laboratory, Guangzhou Chest Hospital, Guangzhou, China;2. Katharine Hsu International Research Center of Human Infectious Diseases, Shandong Provincial Chest Hospital, Shandong University, Jinan, China;3. Central Laboratory, Chongqing Public Health Medical Center, Chongqing, China;4. Department of Bacteriology and Immunology, Beijing Key Laboratory on Drug-Resistant Tuberculosis Research, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, China;5. Department of Clinical Laboratory, Hunan Chest Hospital, Changsha, China;6. Department of Clinical Laboratory, The Eighth Affiliated Hospital of Xinjiang Medical University, Urumqi, China;7. Department of Clinical Laboratory, Anhui Chest Hospital, Hefei, China;8. Department of Clinical Laboratory, Changsha Central Hospital, Changsha, China;9. Department of Clinical Laboratory, Affiliated Infectious Disease Hospital of Nanning, Guangxi Medical University, Nanning, China;10. Department of Clinical Laboratory, Shenyang Tenth People''s Hospital, Shenyang, China;11. Department of Clinical Laboratory, Xi''an Chest Hospital, Xi''an, China;12. Department of Clinical Laboratory, Guangxi Zhuang Autonomous Region Longtan Hospital, Liuzhou, China;13. Department of Clinical Laboratory, Henan Provincial Infectious Diseases Hospital, Zhengzhou, China;14. Department of Clinical Laboratory, The Fifth People''s Hospital of Suzhou, Suzhou, China;15. Department of Clinical Laboratory, The Third People''s Hospital of Kunming, Kunming, China;p. Department of Clinical Laboratory, Harbin Chest Hospital, Harbin, China;q. Department of Clinical Laboratory, The First Affiliated Hospital of Xiamen University, Xiamen, China;r. Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, China;s. Clinical Center on Tuberculosis Control, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, China;t. Department of Laboratory Quality Control, Innovation Alliance on Tuberculosis Diagnosis and Treatment (Beijing), Beijing, China;1. Human Studies Facility, US Environmental Protection Agency, 104 Mason Farm Road, Chapel Hill, NC 27514, USA;2. Oak Ridge Institute for Science and Education, Oak Ridge, TN, USA;3. University of North Carolina, Chapel Hill, NC, USA;4. Innovate! Inc., Alexandria, VA, USA;5. Department of Health and Human Services, Raleigh, NC, USA;6. Duke University Medical Center, Durham, NC, USA |
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| Abstract: | Identification of Mycobacterium species is difficult due to a complex and rapidly changing taxonomy, the failure of 16S rRNA to discriminate many closely related species and the unreliability of phenotypic testing. We investigated a collection of nontuberculous mycobacteria (NTM) strains isolated from suspected tuberculosis patients at Tuberculosis Reference Centre (Ahvaz, Iran) and Masoud Laboratory (Tehran, Iran) during 2008–2012 to evaluate the species spectrum of NTM isolates.Based on phenotypic tests, the isolates were identified up to species or complex level; however they were heterogonous by hsp65-PCR restriction fragment length polymorphism analysis (PRA) method. Representative isolates from each hsp65-PRA pattern, were subjected to identification using single locus and multi locus sequence analysis (MLSA) based on 16S rRNA, rpoB, hsp65 and 16S–23S internal transcribes spacer (ITS) fragments to determine their taxonomic affiliations.All 92 NTM isolates from different clinical specimens were considered as etiological agents causing disease according to American Thoracic Society (ATS) guideline. Phenotypic evaluation alone assigned 66 (72%) isolates to a species or complex level and consequently 76 (82%) isolates showed previously reported hsp65-PRA patterns. Although sequence base identification using single locus such as 16S rRNA, rpoB, hsp65 or ITS identified the isolates up to species level, MLSA correctly identified 16 different species of NTM from clinical isolates. In summary, four-locus MLSA is a reliable method for elucidating taxonomic data and reliable species identification of Mycobacterium isolates and therefore, would be more feasible for routine use in Tuberculosis (TB) reference laboratory. |
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| Keywords: | Infection Identification Multi locus sequence analysis (MLSA) |
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