Evaluation of the BACTEC MGIT 960 in comparison with BACTEC 460 TB for detection and recovery of mycobacteria from clinical specimens.

The BACTEC MGIT system (M960), a fully automated, non radiometric instrument, designed for rapid detection of acid fast bacilli (AFB) from clinical specimens, was compared with the radiometric BACTEC 460 system (B460) and Löwenstein-Jensen (L-J) solid medium. A total of 1,093 respiratory and extrapulmonary specimens were decontaminated by the NALC-NaOH standard method, and randomly inoculated into the media. A total of 122 mycobacteria were recovered, including 47 Mycobacterium tuberculosis complex (MTB) isolates and 75 nontuberculous mycobacteria (NTM) isolates. Overall recovery rates were 59% for M960 system (p < 0.0001), 58.2% for L-J. medium (p < 0.0001) and 82% for Bactec 460 system, whereas rates for MTB alone were 91.5, 76.6 (p = 0.007), and 95.7%, respectively. The combination of M960 or B460 systems with L-J medium showed the same recovery rates for MTB strains (97.9%), whereas NTM rates were 68% (p < 0.0001) and 93.3%, respectively. Mean time to detection of smear-positive MTB, smear-negative MTB, and NTM were 12.2, 13.4, and 23.3 days, respectively, with the M960, 11.7, 21.3, and 24.8 days with the B460 and 20.4, 28.7, and 28.4 days with L-J medium. The M960 system showed a contamination rate of 9.8%, while B460 and L-J medium showed contamination rates of 4.3 and 3.8% respectively. In conclusion, the M960 system appeared to be accurate and rapid for the recovery of MTB, but reduced recovery of NTM and a high number of contaminated cultures deserve further study in order to assess if this system can represent a valuable alternative to the radiometric system.

Performance assessment of two commercial amplification assays for direct detection of Mycobacterium tuberculosis complex from respiratory and extrapulmonary specimens.

The new BDProbeTec ET Mycobacterium tuberculosis Complex Direct Detection Assay (DTB) was compared with the enhanced M. tuberculosis Amplified Direct Test (AMTDII). The system is an automated walkaway system characterized by simultaneous DNA amplification (strand displacement amplification) and real-time fluorometric detection. It also contains an internal amplification control (IAC) designed to identify inhibition from the processed samples. The AMTDII assay amplifies rRNA by transcription-mediated amplification; it uses hybridization with a chemoluminescent probe as a detection system and is entirely manual. A total of 515 N-acetyl-L-cysteine-sodium hydroxide-decontaminated respiratory (n = 331) and extrapulmonary (n = 184) sediments (from 402 patients) were tested in parallel by both assays. The results were compared with those of acid-fast staining and culture (solid plus liquid media), setting the combination of culture and clinical diagnosis as the "gold standard." Culture results from the tested specimens were as follows: 121 Mycobacterium tuberculosis complex (MTB) (98 smear-positive), 46 nontuberculous mycobacteria (38 smear-positive), and 338 culture-negative results. After resolution of the discrepant results, the percent sensitivity, percent specificity, and positive and negative likelihood ratios for AMTDII were 88%, 99.2%, 110, and 0.11 for respiratory specimens and 74.3%, 100%, 740, and 0.26 for extrapulmonary specimens, respectively. The corresponding values for DTB were 94.5%, 99.6%, 235, and 0.05 for respiratory specimens and 92.3%, 100%, 920, and 0.07 for extrapulmonary specimens, respectively. The cumulative difference for all tuberculosis-positive extrapulmonary specimens was significant (P = 0.03). The overall inhibition rate for DTB was 5% (26 specimens). We conclude that both amplification assays proved to be rapid and specific for the detection of MTB in clinical samples and particularly feasible for a routine laboratory work flow. DTB combines a labor-intensive specimen preparation procedure with a completely automated amplification and detection. Finally, differences between AMTDII and DTB sensitivities were associated with the presence of inhibitory samples that the former assay, lacking IAC, could not detect.