Prevention of false resistance results obtained in testing the susceptibility of Mycobacterium tuberculosis to pyrazinamide with the Bactec MGIT 960 system using a reduced inoculum.

The susceptibility of 211 clinical isolates of Mycobacterium tuberculosis complex (201 M. tuberculosis and 10 Mycobacterium bovis isolates) to pyrazinamide (PZA) was assessed by the nonradiometric Bactec MGIT 960 system (M960). Detection of PZA resistance was followed by a repeat testing using a reduced inoculum (RI) of 0.25 ml instead of 0.5 ml. According to the first M960 analysis, resistance was observed in 55 samples. In the RI assay, 32 samples turned out to be susceptible and 23 proved to be resistant (58.2% false positivity). The Bactec 460 assay confirmed as resistant those strains detected by the RI assay, while discrepant results were found susceptible. Mutation analysis performed on 13 M. tuberculosis isolates detected pncA mutations in 11 samples. On the basis of our data, we suggest using the RI assay to confirm all PZA resistance results obtained with the standard M960 assay. Further studies are required to confirm our findings.

Unreliable detection of Mycobacterium xenopi by the nonradiometric Bactec MGIT 960 culture system.

From June 2006 to December 2007, 3,648 clinical specimens consecutively received for mycobacterial culture were investigated. Each processed sample was inoculated into Bactec MGIT 960 liquid medium and a Löwenstein-Jensen slant. Tubes that were flagged as positive by the instrument as well as those determined to be negative after 42 days of incubation were removed, visually inspected for growth, and checked for the presence of acid-fast bacilli. Three hundred sixty-nine mycobacterial strains were recovered; of the 44 Mycobacterium xenopi isolates recovered by MGIT medium, only 13 were detected by the instrument (P<0.0001). Most tubes yielding M. xenopi exhibited a peculiar pattern of growth characterized by a scant number of round, yellow-pigmented granules instead of the fine, evenly dispersed clumps usually observed for mycobacteria. It is suggested to check all individual tubes discarded by the MGIT 960 system at the end of the incubation period to prevent a significant amount of previously undetected growth from being missed.

Mycobacterium celatum pulmonary infection in the immunocompetent: case report and review.

Mycobacterium celatum has been shown to cause disease in immunocompromised patients. We report a case of serious pulmonary infection caused by M. celatum in an apparently immunocompetent patient and review the characteristics of two other reported cases. Clinical and radiologic symptoms and signs included cough, malaise, and weight loss associated with cavitary lesions and pulmonary infiltrates. Although M. celatum is easy to detect in clinical specimens by liquid and solid media, it may be misidentified as a member of the M. tuberculosis complex or as M. xenopi. M. celatum pulmonary infection appears to respond to antimycobacterial chemotherapy, particularly with clarithromycin.

Mycobacterium lentiflavum as an emerging causative agent of cervical lymphadenitis.

A lymph node excision was performed on a 45-year-old woman with left cervical swelling. The disorder which developed after the patient had undergone oral surgery for a severe periodontal disease failed to respond to antimicrobial chemotherapy. A mycobacterial strain subsequently identified by high-performance liquid chromatography analysis of cell wall mycolic acids as Mycobacterium lentiflavum grew from the excised specimen. This case and previously published reports highlight the relevance of M. lentiflavum as an emerging causative agent of mycobacterial cervical lymphadenitis.

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.