Comparison of the BACTEC MGIT 960 and BACTEC 460TB systems for detection of mycobacteria in clinical specimens

The reliability of the Mycobacteria Growth Indicator Tube (MGIT) 960 system for rapid detection of mycobacteria in clinical specimens was evaluated and compared to the radiometric method (BACTEC 460TB) and to mycobacterial culture on Lowenstein-Jensen (LJ) medium. Clinical specimens (n = 590) were tested without selection. A total of 121 (20.5%) isolates of mycobacteria were recovered; 98 (81.0%) of them were recovered with the BACTEC 460TB system, 86 (71.1%) were recovered with the BACTEC MGIT 960 system, and 55 (45.5%) were recovered with LJ medium (MGIT 960 versus BACTEC 640TB, p >0.05; MGIT 960 or BACTEC 460TB versus LJ, p <0.001). The mean time to detection (TTD) was 18 da for BACTEC 460 TB, and 13 da for BACTEC MGIT 960. The mean time to detection in each system, based upon data where both systems were culture positive, was significantly different (16.6 da for BACTEC 460TB and 13 da for BACTEC MGIT 960, p<0.001). The contamination rate of the BACTEC MGIT 960 system was 13.2%, which was intermediate between the BACTEC 460TB system (11.7%) and the LJ medium (14.7%). These data indicate that the fully automated MGIT 960 system is an accurate, non-radiometric alternative to the BACTEC 460TB method for rapid detection of mycobacteria in a clinical setting.     

Meta-analysis of BACTEC MGIT 960 and BACTEC 460 TB, with or without solid media, for detection of mycobacteria

In a meta-analysis of 10 studies, the BACTEC 960/MGIT and BACTEC 460 systems showed a sensitivity and specificity in detecting mycobacteria (1,381 strains from 14,745 clinical specimens) of 81.5 and 99.6% and 85.8 and 99.9%, respectively. Combined with solid media, the sensitivity of the two systems increased to 87.7 and 89.7%, respectively.     

Comparison of BACTEC MGIT 960 system and BACTEC 460 TB system for growth and detection of Mycobacteria from clinical specimens

This study was carried out to compare the performance of BACTEC MGIT 960 with the BACTEC 460 TB for growth and detection of Mycobacteria from human clinical specimens. The BACTEC MGIT 960 instrument is a fully automated system that utilizes MGIT tubes containing an oxygen sensor embedded in silicon at the bottom and filled with 7 mL of modified Middlebrook 7H9 broth. Identical samples were inoculated into the two automated systems and incubated for six weeks. Over a period of three months, 279 specimens were decontaminated and processed according to the standard CDC NALC/NaOH method, using the commercial MycoPrep kit. Forty-two specimens (15%) yielded Mycobacterium tuberculosis; 37 (88%) were detected by the fluorescent BACTEC MGIT 960 and 35 (83%) detected by the radiometric BACTEC 460 TB. Fifteen specimens (5%) yielded Mycobacterium Other Than Tuberculosis (MOTT); 10 (66%) were detected by BACTEC MGIT 960 and 11 (73%) detected by BACTEC 460 TB. The average time to detection and contamination rates and the average time to obtain results of antimicrobial susceptibility tests between the two systems were compared. The performance of the BACTEC MGIT 960 was comparable to the BACTEC 460 TB system which has been the "Gold Standard" for automated detection of TB. The former was more rapid, as sensitive and less labour intensive than the BACTEC 460. Our data demonstrates that the BACTEC MGIT 960 system is an accurate, automated and a non-radioactive alternative to the BACTEC 460 TB for the culture and susceptibility testing of M. tuberculosis.     

Multicenter Evaluation of the BACTEC MGIT 960 System for Recovery of Mycobacteria

We evaluated the BACTEC MGIT 960 system, which is a fully automated, noninvasive system for the growth and detection of mycobacteria with a capacity to incubate and continuously monitor 960 7-ml culture tubes. We studied 3,330 specimens, 2,210 respiratory and 1,120 nonrespiratory specimens, collected from 2,346 patients treated at six sites. Processed specimens were inoculated into the BACTEC MGIT 960 and BACTEC 460 TB systems, as well as onto Lowenstein-Jensen slants and Middlebrook 7H11/7H11 selective plates. From all culture systems, a total of 362 isolates of mycobacteria were recovered; these were recovered from 353 specimens collected from 247 patients. The greatest number of isolates of mycobacteria (289, or 80% of the 362 isolates) was recovered with the BACTEC MGIT 960, followed by the BACTEC 460 TB (271, or 75%) and solid media (250, or 69%). From all culture systems a total of 132 isolates of Mycobacterium tuberculosis complex were recovered. The greatest number of isolates of M. tuberculosis complex was recovered when liquid medium was combined with conventional solid media; the number recovered with BACTEC 460 TB plus solid media was 128 (97%), that recovered with BACTEC MGIT 960 plus solid media was 121 (92%), that recovered with BACTEC 460 TB was 119 (90%) and that recovered with all solid media combined was 105 (79%). The recovery with BACTEC MGIT 960 alone was 102 (77%). The mean times to detection (TTD) for M. tuberculosis complex were 14.4 days for BACTEC MGIT, 15.2 days for BACTEC 460 TB, and 24.1 days for solid media. The numbers of isolates of Mycobacterium avium complex (MAC) recovered were 172 (100%) for all systems, 147 (85%) for BACTEC MGIT 960, 123 (72%) for BACTEC 460 TB, and 106 (62%) for all solid media combined. The TTD for MAC in each system were 10.0 days for BACTEC MGIT 960, 10.4 days for BACTEC 460 TB, and 25.9 days for solid media. Breakthrough contamination rates (percentages of isolates) for each of the systems were 8.1% for BACTEC MGIT 960, 4.9% for BACTEC 460 TB, and 21.1% for all solid media combined.     

Xpert Mtb/RIF检测与BACTEC MGIT960结核培养药敏联合检测在结核病诊治中的应用分析

目的评估Xpert Mtb/RIF检测与BACTEC MGIT 960结核培养药敏联合检测在结核病诊治中的应用价值。方法对临床结核病患者的标本进行Xpert Mtb/RIF检测,同时进行BACTEC MGIT 960结核培养、鉴定、药敏试验。以BACTEC MGIT 960结核培养及药敏试验为金标准,分析Xpert Mtb/RIF检测的敏感性和特异性。结果以BACTEC MGIT 960结核培养及药敏试验为金标准,Xpert Mtb/RIF检测结核分枝杆菌的敏感性为87.7%(100/114),Xpert Mtb/RIF检测利福平耐药的敏感性为90%(45/50)、特异性为91.3%(53/58)。结论 Xpert Mtb/RIF检测敏感性及特异性高, BACTEC MGIT 960结核培养及药敏试验联合应用有助于及早进行有效诊治。     

Comparison of the BACTEC MGIT 960 and ESP culture system II for growth and detection of mycobacteria

The performances of two continuously monitoring mycobacterial culture systems-ESP Culture System II (ESP II; Trek Diagnostics, Inc. , Westlake, Ohio) and BACTEC MGIT 960 (BD Biosciences, Sparks, Md. )-were compared. In addition to both liquid media, all specimens were plated onto Middlebrook 7H11/7H11 selective agar. A total of 3, 151 specimens of all types (56.3% were respiratory specimens) were cultured; 231 (7.3%) yielded mycobacteria. The most common species recovered were Mycobacterium avium complex (69 isolates) and Mycobacterium tuberculosis complex (MTBC; 65 isolates). The recovery rates for ESP II, BACTEC MGIT 960, and Middlebrook agar, respectively, were 71.2, 63.9, and 61.8% for all mycobacteria; 70.2, 72.6, and 66.3% for all mycobacteria except Mycobacterium gordonae; and 73.8, 84.6, and 87.7% for MTBC. For liquid plus solid medium combinations, recovery rates for all mycobacteria and for MTBC, respectively, were 84.1 and 92.3% for ESP II plus Middlebrook agar and 81.5 and 98.5% for BACTEC MGIT 960 plus Middlebrook agar. The differences in recovery of all mycobacteria by ESP II and by BACTEC MGIT 960 were not significant; for the individual species, the only significant difference was recovery of more isolates of M. gordonae by ESP II. For those isolates recovered in both automated systems, mean times to detection of all mycobacteria and MTBC, respectively, were 15.8 and 17.4 days for ESP II and 12.5 and 11.9 days for BACTEC MGIT 960 (P < 0.05). False-positive signals occurred with 23 (0.7%) BACTEC MGIT 960 cultures and 84 (2.7%) ESP II cultures (P < 0.01). Overall contamination rates were 17.1% for BACTEC MGIT 960, 18.9% for ESP II, and 11.0% for Middlebrook agar. In summary, the ESP II and BACTEC MGIT 960 systems performed comparably with regard to growth and detection of mycobacteria, and the contamination rates were similar. However, with ESP II, times to detection of all mycobacteria and of MTBC were significantly longer, the recovery rate of M. gordonae was significantly higher, and the number of false-positive signals was greater than with BACTEC MGIT 960.     

A comparative study for the detection of Mycobacteria by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination

PURPOSE: To compare BACTEC MGIT 960 (M960) with conventional culture on Lowenstein Jensen (LJ) media and direct acid fast bacilli (AFB) smear examination for the detection of Mycobacteria in clinical samples obtained from suspected cases of pulmonary and extra pulmonary tuberculosis (TB). METHODS: A total of 500 samples were processed for direct AFB smear examination, and culture on M960 and LJ media. RESULTS: Two hundred fifty-eight out of 500 (51.6%) isolates of Mycobacteria were obtained by combined use of the two culture methods. Two hundred and fifty-three (50.6%) were positive in culture by M960 and LJ media and 28% (140/500) by direct AFB smear examination. The positivity rate of M960 system alone was 34.10% (88/258) and of LJ alone was 1.93% (5/258). Average time to detect growth (TTD) was 9.66 days by M960 and 28.81 days by LJ. CONCLUSIONS: M960 system is a rapid and sensitive method for early diagnosis of pulmonary and extrapulmonary TB. But for maximum recovery of Mycobacteria , a combination of both M960 and LJ media should be used.     

Testing of susceptibility of Mycobacterium tuberculosis to pyrazinamide with the nonradiometric BACTEC MGIT 960 system

The reliability of the novel BACTEC MGIT 960 pyrazinamide (PZA) kit (Becton Dickinson Microbiology Systems, Sparks, Md.) was assessed for testing of susceptibility of Mycobacterium tuberculosis to PZA. Results generated by the BACTEC MGIT 960 system (Becton Dickinson) were compared with those obtained with the BACTEC 460TB system. Extensive proficiency testing (phase I) and reproducibility testing (phase II) as well as susceptibility testing of blinded strains of M. tuberculosis from the Centers for Disease Control and Prevention (phase III) were performed prior to testing 58 strains isolated from clinical specimens (phase IV). After resolution of discrepant results obtained by the two BACTEC methods by two other laboratories which acted as independent arbiters (phase V), overall agreement of the BACTEC MGIT 960 system with the BACTEC 460TB system for PZA testing of phase IV strains was 96.6%. Between the two systems there was no statistically significant difference in time until results were obtained, i.e., 6.8 days (BACTEC MGIT 960) versus 5.4 days (BACTEC 460TB), the latter not counting the time required for a subculture with a growth index of 200, however. The new BACTEC MGIT PZA susceptibility testing procedure works equally well for inocula prepared from liquid (MGIT) and solid (L?wenstein-Jensen) cultures. PZA MGIT medium in plastic tubes yielded results equivalent to medium dispensed in glass tubes.     

Use of BACTEC MGIT 960 for Recovery of Mycobacteria from Clinical Specimens: Multicenter Study

The BACTEC MGIT 960 instrument is a fully automated system that exploits the fluorescence of an oxygen sensor to detect growth of mycobacteria in culture. Its performance was compared to those of the radiometric BACTEC 460 instrument and egg-based Lowenstein-Jensen medium. An identical volume of sample was inoculated in different media, and incubation was carried out for 6 weeks with the automatic systems and for 8 weeks on solid media. A total of 2,567 specimens obtained from 1,631 patients were cultured in parallel. Mycobacteria belonging to nine different taxa were isolated by at least one of the culture systems, with 75% of them being represented by Mycobacterium tuberculosis complex. The best yield was obtained with the BACTEC 460 system, with 201 isolates, in comparison with 190 isolates with the BACTEC MGIT 960 system and 168 isolates with Lowenstein-Jensen medium. A similar but not significant difference was obtained when the most-represented organisms, the M. tuberculosis complex, Mycobacterium xenopi, and the Mycobacterium avium complex, were analyzed separately and when combinations of a solid medium with the BACTEC MGIT 960 system and with the BACTEC 460 system were considered. The shortest times to detection were obtained with the BACTEC MGIT 960 system (13.3 days); 1.5 days earlier than that with the BACTEC 460 system (14.8 days) and 12 days earlier than that with Lowenstein-Jensen medium (25.6 days). The BACTEC MGIT 960 system had a contamination rate of 10.0%, intermediate between those of the radiometric system (3.7%) and the egg-based medium (17.0%). We conclude, therefore, that the BACTEC MGIT 960 system is a fully automated, nonradiometric instrument that is suitable for the detection of growth of tuberculous and other mycobacterial species and that is characterized by detection times that are even shorter than that of the "gold standard," the BACTEC 460 system. The contamination rate was higher than that for the radiometric BACTEC 460 system and needs to be improved.     

Presumptive identification of Mycobacterium tuberculosis complex based on cord formation in BACTEC MGIT 960 medium

We considered samples received for culture of mycobacteria using BACTEC MGIT 960 system over a period of 1 year. Tubes flagged positive by MGIT were evaluated for presence of serpentine cording. The cord formation was compared with isolates identified as Mycobacterium tuberculosis complex (MTC) based on p-nitrobenzoic acid (PNB) test. Cords were found in 591 isolates of which 584 (98.8%) were confirmed as MTC. The sensitivity and specificity of cord formation were found to be 99.7% and 89.9%, respectively.     

Rapid and reliable method for quantification of Mycobacterium paratuberculosis by use of the BACTEC MGIT 960 system

A simple method for the enumeration of viable Mycobacterium paratuberculosis cells was developed and evaluated using the MGIT 960 culture system. For each of 12 M. paratuberculosis strains isolated from either cattle or humans, single-cell suspensions of M. paratuberculosis cells were adjusted to an optical density at 600 nm of 1.00 (10(7.6) to 10(8.2) cells/ml), and serial dilutions were prepared. Standard curves were established by relating the MGIT time-to-detection data to the log10 CFU for these suspensions using standard plate counting and BACTEC 460 results as reference methods. Universal and strain-specific standard quantification curves were generated. A one-phase exponential decay equation best fit the universal standard curve and strain-specific curves (R2 of 0.96 and >0.99, respectively). Two subgroups within the universal curves were distinguished: one for laboratory-adapted strains and the other for recently isolated low-passage bovine strains. The predictive errors for log(10) estimations using the universal standard curve, each subgroup's standard curve, and strain-specific curves were +/-0.87, +/-0.45, and +/-0.31 log10 units, respectively. CFU estimations by all three standard curves were highly reproducible, regardless of the M. paratuberculosis strain or inoculum volume. In comparison with the previously described BACTEC 460 M. paratuberculosis counting method, quantification with MGIT 960 was less expensive, more rapid, more accurate, and more sensitive (<10 CFU). This MGIT counting method has broad applications for studies requiring the quantification of viable M. paratuberculosis cells, such as drug susceptibility testing or environmental survival studies.     

Growth detection failures by the nonradiometric Bactec MGIT 960 mycobacterial culture system

Mycobacterial growth in liquid culture can go undetected by automated, nonradiometric growth detection systems. In our laboratory, instrument-negative tubes from the Bactec MGIT 960 system are inspected visually for clumps suggestive of mycobacterial growth, which (if present) are examined by acid-fast smear analysis. A 3-year review demonstrated that ～1% of instrument-negative MGIT cultures contained mycobacterial growth and that 10% of all cultures yielding mycobacteria were instrument negative. Isolates from instrument-negative MGIT cultures included both tuberculous and nontuberculous mycobacteria.     

Evaluation of the invader assay with the BACTEC MGIT 960 system for prompt isolation and identification of mycobacterial species from clinical specimens

Rapid and accurate identification of mycobacterial species is essential for patient management. We describe the use of the Invader assay in conjunction with the BACTEC MGIT 960 system that together provide an efficient procedure for clinical use. This assay discriminates single-base differences (e.g., genotyping single-nucleotide polymorphisms) under homogeneous and isothermal conditions and can measure directly on genomic DNA without prior target DNA amplification. To identify a wide variety of mycobacterial species, 20 Invader probes were designed to target the 16S rRNA gene and the 16S-23S rRNA gene internal transcribed spacer 1 (ITS-1) region. To validate the Invader probes, we used 78 ATCC strains, and 607 clinical mycobacterial strains, which were identified by DNA sequencing of the 16S rRNA gene and ITS-1. The Invader assay could accurately identify and differentiate these strains according to target sequences. Moreover, it could detect and identify 116 (95.1%) of 122 positive liquid cultures from the BACTEC MGIT 960 system and did not react to 83 contaminated MGIT cultures. Species identification takes 6.5 h by the Invader assay: 2.0 h for DNA extraction, 0.5 h for handling, and up to 4 h for the Invader reaction. The Invader assay has the speed, ease of use, and accuracy to be an effective procedure for the bacteriological diagnosis of mycobacterial infections.     

Usefulness of the BACTEC MGIT 960 system for isolation of mycobacterium tuberculosis from sputa subjected to long-term storage

The recovery of Mycobacterium tuberculosis from sputa positive or negative for acid-fast bacilli that were stored for 17 +/- 7 days and inoculated in the BACTEC MGIT 960 system (MGIT) was higher than that from sputa inoculated in Lowenstein-Jensen medium. MGIT is useful for isolation of M. tuberculosis from sputa subjected to long-term storage.     

Improved detection of Mycobacterium spp. using the Bactec MGIT 960 system

Until 1987, the notification rate for mycobacterial infection was on the decline; however, it now appears to be increasing once more. The reason for this may be multifactoral and include improved reporting of diagnosed cases, increased infection of an ageing population, homelessness, immunosuppression (e.g. due to human immunodeficiency virus infection), and immigration of people from countries where tuberculosis is endemic. This rising incidence and the increasing importance of resistant organisms mean that rapid identification by the clinical microbiology laboratory is required, and this is where an automated detection system can be an advantage. Over a two-year period, 2743 clinical specimen were examined for Mycobacterium spp. using the Bactec MGIT 960, and 286 were positive. Time to detection ranged from three to 14 days (mean: 9.3 days), and M. tuberculosis was recovered from 214 (75.5%). Contamination rate was higher (8.6%) than with manual methods, however. On balance, the Bactec MGIT 960 system proved a valuable tool in the routine microbiology laboratory.     

Early detection of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> complex in BACTEC MGIT cultures using nucleic acid amplification

We evaluated the application of nucleic acid amplification (NAA) in liquid cultures for the early detection of Mycobacterium tuberculosis. The Cobas TaqMan MTB test, IS6110 real-time PCR, and hsp65 PCR-restriction fragment length polymorphism (RFLP) analysis were used to detect BACTEC MGIT 960 (MGIT) cultures on days 3, 5, 7, and 14. The procedure was initially tested with a reference strain, H37Rv (ATCC 27294). Subsequently, 200 clinical specimens, including 150 Acid Fast bacillus (AFB) smear-positive and 50 AFB smear-negative samples, were examined. The Cobas TaqMan MTB test and IS6110-based PCR analysis were able to detect M. tuberculosis after 1 day when the inoculum of H37Rv was >3 x 10^?2 CFU/ml. After a 5-day incubation in the MGIT system, all three NAA assays had a positive detection regardless of the inoculum size. After a 1-day incubation of the clinical specimens in the MGIT system, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the Cobas TaqMan MTB assay were 70.2%, 100%, 100%, and 82.3% respectively. For IS6110-based PCR analysis, these values were 63.1%, 100%, 100%, and 78.9%, and were 88.1%, 100%, 100%, and 92.1% respectively for hsp65 PCR-RFLP analysis. After a 3-day incubation, the specificity and PPV were 100% for all three NAA tests; the Cobas TaqMan MTB assay had the best sensitivity (97.6%) and NPV (98.3%). The sensitivity, specificity, PPV, and NPV for conventional culture analysis were 98.8%, 100%, 100%, and 99.1%. Thus, NAA may be useful for the early detection of M. tuberculosis after 3 days in MGIT.     

Multicenter laboratory evaluation of the MB/BacT Mycobacterium detection system and the BACTEC MGIT 960 system in comparison with the BACTEC 460TB system for susceptibility testing of Mycobacterium tuberculosis

In this multicenter study, the reliability of two nonradiometric, fully automated systems, the MB/BacT and BACTEC MGIT 960 systems, for testing the susceptibilities of 82 Mycobacterium tuberculosis strains to isoniazid, rifampin, ethambutol, and streptomycin was evaluated in comparison with the radiometric BACTEC 460TB system. The arbitration of discrepant results was done by the reanalysis of the strain, the determination of the MIC, and the molecular characterization of some resistance determinants. The overall level of agreement with BACTEC 460TB results was 96% with the MB/BacT test and 97.2% with the BACTEC MGIT 960 system. With both methods, the level of agreement with BACTEC 460TB results was 96.3% for isoniazid, 98.8% for rifampin, and 98.8% for ethambutol. The level of agreement for streptomycin was 90.2% with MB/BacT and 97.5% with BACTEC MGIT 960. Overall, there were 11 very major errors and 2 major errors with the MB/BacT method and 5 very major errors and 2 major errors with the BACTEC MGIT 960 system. In general, the MB/BacT and BACTEC MGIT 960 systems showed good performance for susceptibility testing with first-line antituberculosis drugs.     

Evaluation of BACTEC MGIT 960 and BACTEC 460TB systems for recovery of mycobacteria from clinical specimens of a university hospital with low incidence of tuberculosis

Clinical samples obtained over a period of 8 months (n = 2,624) were processed in parallel with the BACTEC 460TB system, with the MGIT 960 system, and in L?wenstein-Jensen (LJ) medium, resulting in the recovery of 127 mycobacteria. Recovery rates in combinations of the BACTEC 460TB or MGIT 960 system with LJ were, respectively, 94.7 and 94.7% for Mycobacterium tuberculosis complex (n = 57) and 91.4 and 70.0% for nontuberculous mycobacteria (n = 70). Contamination rates, elevated in the MGIT 960 system, were associated with patients (cystic fibrosis) and type of material but not with transport time. Detection time was reduced in the MGIT 960 system.     

Evaluation of BACTEC Mycobacteria Growth Indicator Tube (MGIT 960) Automated System for Drug Susceptibility Testing of Mycobacterium tuberculosis

The reliability of the BACTEC MGIT 960 system, an automated version of the Mycobacteria Growth Indicator Tube (MGIT), for antimicrobial susceptibility testing of Mycobacterium tuberculosis was evaluated on 78 clinical isolates. Rifampin (RMP), isoniazid (INH), streptomycin (SM), and ethambutol (EMB) were tested at the following concentrations: 1.0 microg/ml for RMP, 0.1 and 0.4 microg/ml for INH, 1.0 and 4.0 microg/ml for SM, and 5.0 and 7.5 microg/ml for EMB. Results were compared with those obtained by the BACTEC 460 TB radiometric system. Initially the reproducibility study showed 99.5% agreement on repeat testing with all the four drugs. With susceptibility testing of clinical isolates, excellent agreement between the two systems was found for all the drugs. A total of nine major errors were observed for only three isolates, resistant according to BACTEC MGIT 960 and susceptible according to BACTEC 460 TB, to SM (4.0 microg/ml), INH (0.1 microg/ml), and EMB (5.0 microg/ml) (one isolate) and to SM (1.0 microg/ml), INH (0.4 microg/ml), and EMB (5.0 microg/ml) (two isolates). When these isolates were tested by using the conventional proportion method on L?wenstein-Jensen medium, agreement with BACTEC MGIT 960 was found for five results and with BACTEC 460 TB for the remainder. The time to report results was 7.9 days by MGIT 960 and 7.3 days by BACTEC 460 TB, which was not found statistically significant (P > 0.05). In conclusion, the performance of BACTEC MGIT 960 was found similar to that of BACTEC 460 TB and this new system can be considered a good alternative to the radiometric method for routine susceptibility testing of M. tuberculosis.