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.

Validation of the agar proportion and 2 liquid systems for testing the susceptibility of Mycobacterium tuberculosis to moxifloxacin.

Moxifloxacin (MOX), an 8-methoxyquinolone compound, is now widely used for the treatment of bacterial infections and also accepted as 2nd-line drug for the treatment of multidrug-resistant (MDR) tuberculosis. To tentatively correlate the clinical outcome with in vitro results, we sought to set up susceptibility test conditions for Mycobacterium tuberculosis against MOX by using the reference agar proportion method, the BACTEC 460 radiometric system, and the recently validated nonradiometric BACTEC MGIT 960 system. Our aim was to determine the critical MOX test concentration to be used with the abovementioned methods for routine susceptibility testing. MICs were determined for 20 pan-susceptible strains, 10 MDR strains, and 10 fluoroquinolone-resistant strains with defined gyrA mutations. MOX MICs resulted in a bimodal pattern with values for gyrA mutants considerably higher than those for pan-susceptible and MDR strains. Our data showed that a concentration of 0.5 microg/mL allowed a clear-cut separation between susceptible and resistant strains when tested by all the studied methods. Confirmatory test with a subset of pan-susceptible and MDR isolates appeared to validate the selected critical concentration. The MOX-resistant strains were almost isolated from patients with prior fluoroquinolone exposure.

Clinical suspicion as a primary guidance to use commercial amplification tests for rapid diagnosis of pulmonary tuberculosis.

The Abbott LCx (Abbott Park, IL) Mycobacterium tuberculosis complex is a commercial amplification assay discontinued from the European market in 2002. A prospective clinical study was carried out to evaluate the clinical utility of the above test as applied by specialists for the rapid diagnosis of active pulmonary tuberculosis (PTB). According to the physician's clinical judgment, patients were classified into 3 groups (low, intermediate, and high) aiming to estimate the probability of active disease. The gold standard for final diagnosis was based on microbiologic and clinical information including data from a 6-month follow-up period. Sensitivities and specificities of rapid microbiologic tests were compared with those based on an integrated approach including clinical evaluation plus the above tests. The incidence of PTB in 214 patients was 13.1%. The basis for initial treatment of PTB was smear-positive results in 46%, positive LCx results in 29%, and clinical suspicion in 18%. For the remaining 7%, therapy was started upon receipt of culture results. The sensitivity, specificity, and positive and negative predictive values of the LCx assay were 68%, 99%, 95%, and 95%, respectively. In comparison, they were 93%, 99%, 96%, and 99%, respectively, for the combination of clinical evaluation plus the LCx test. It is concluded that in patients with high-to-moderate pretest probabilities, the combination of clinical judgment and amplification results strongly enhances a rapid and correct diagnosis of PTB.

Mycobacterium triplex pulmonary disease in immunocompetent host.

Mycobacterium triplex, a recently described, potentially pathogenic species, caused disease primarily in immunocompromised patients. We report a case of pulmonary infection due to this mycobacterium in an immunocompetent patient and review the characteristics of two other cases. In our experience, Mycobacterium triplex pulmonary infection is unresponsive to antimycobacterial chemotherapy.

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.

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.

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.