Identification of non-tuberculous mycobacteria: utility of the GenoType Mycobacterium CM/AS assay compared with HPLC and 16S rRNA gene sequencing

• Published in: Journal of medical microbiology Vol. 58; no. 7; pp. 900 - 904
• Main Authors: Lee, Andie S, Jelfs, Peter, Sintchenko, Vitali, Gilbert, Gwendolyn L
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.07.2009; Society for General Microbiology
• Subjects: Bacterial diseases; Bacteriological Techniques; Bacteriology; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; Genotype; Human bacterial diseases; Infectious diseases; Medical sciences; Microbiology; Miscellaneous; Mycobacterium - classification; Mycobacterium - isolation & purification; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; Sensitivity and Specificity; Tuberculosis and atypical mycobacterial infections; Mycobacterium; Microbiology; Ribosomal DNA; HPLC chromatography; Genotype; Identification; Mycobacterial infection; Infection; Tuberculosis; Mycobacteriales; Bacteriosis; Mycobacteriaceae; 16S-RNA; Bacteria; Actinomycetes
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.007484-0

1 Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Sydney, Australia 2 Western Clinical School, University of Sydney, Sydney, Australia Correspondence Gwendolyn L. Gilbert l.gilbert{at}usyd.edu.au Received October 22, 2008 Accepted March 27, 2009 Non-tuberculous mycobacteria (NTM) causing clinical disease have become increasingly common and more diverse. A new reverse line probe assay, GenoType Mycobacterium CM/AS (Hain Lifescience), was evaluated for identification of a broad range of NTM. It was compared with phenotypic (HPLC) and molecular (DNA probes, in-house real-time multiplex species-specific PCR, 16S rRNA gene PCR and sequencing) identification techniques, which together provided the reference ‘gold standard’. A total of 131 clinical isolates belonging to 31 Mycobacterium species and 19 controls, including 5 non- Mycobacterium species, was used. Concordant results between the GenoType Mycobacterium assay and the reference identification were obtained in 119/131 clinical isolates (90.8 %). Identification of Mycobacterium abscessus and Mycobacterium lentiflavum by the assay was problematic. The GenoType Mycobacterium assay enables rapid identification of a broad range of potentially clinically significant Mycobacterium species, but some species require further testing to differentiate or confirm ambiguous results. Present address: Infection Control Program, University of Geneva Hospitals, Geneva, Switzerland.