Genotype ntm dr ver 1

Clinical specimens were processed using the sodium hydroxide-N-acetyl-L-cysteine-sodium citrate (NaOH/NALC-Na citrate) method [19 ]. Briefly, clinical specimens were decontaminated and digested with an equal volume of NaOH/NALC-Na citrate digestion solution (2% NaOH, 0.5% NALC, and 1.45% Na-citrate) for 15 min at room temperature. Phosphate buffer (pH 6.8) was added to the processed samples to a final volume of 40 ml, mixed by inversion, and centrifuged at 3,000 xg for 15 min at 12 °C. The supernatant was then discarded, and the sediment was resuspended in a new 2.5 ml phosphate buffer. However, sterile specimens with limited quantity, such as small amounts of cerebrospinal fluid (CSF) and tissue biopsies were processed for culture without digestion-decontamination. A 500 μl of well-mixed sediment was added to the mycobacterial growth indicator tubes (MGIT) (Becton Dickinson, USA) and incubated in the BD BACTEC MGIT 960 System for 6 weeks or until the signal turned positive. In addition, 200 μl of sediment was inoculated on Ogawa solid medium (in-house preparation) for 8 weeks at 37 °C. Mycobacterial identification to species/complex level was achieved by using a line probe assay which was either GenoType MTBC VER 1.X, GenoType Mycobacterium CM VER 2.0, GenoType Mycobacterium AS VER 1.0, or GenoType NTM-DR VER 1.0 (Hain Lifescience, Germany) [20 , 21 , 22 , 23 ].