The BACTEC MGIT 960 culture system (Becton, Dickinson and Company, America) was used for mycobacterial culture. For positive results, strain identification was performed using an Mpb64 monoclonal antibody(Hangzhou Genesis Biodetecton & Biocontrol Ltd, China). First-line drug sensitivity was tested using the BACTEC MGIT 960 system, and second-line drug sensitivity was tested using the absolute concentration method. The drug concentrations for isoniazid, rifampin, levofloxacin, moxifloxacin, amikacin, and capreomycin were 0.1, 1.0, 2.0, 1.0, 30, 40 µg/mL, respectively. The Ziehl-Neelsen acid-fast staining was used for the smear test.
Bactec mgit 960 culture system
Manufactured by BD
Sourced in United States
The BACTEC MGIT 960 culture system is a fully automated mycobacterial detection instrument designed for the rapid and accurate detection of mycobacterial growth in clinical samples. It utilizes a fluorescent monitoring technology to detect the presence of mycobacteria in liquid culture media.
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Lab products found in correlation
6 protocols using bactec mgit 960 culture system
1
Mycobacterial Culture and Drug Sensitivity
2
Genotyping and Whole-Genome Sequencing of MTB
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A total of 40 MTB strains were obtained from the culture collection of the Research Institute of Phtisiopulmonology (St. Petersburg, Russia) and Moscow Scientific-Practical Center of Treatment of Tuberculosis of Moscow Healthcare (Moscow, Russia). The susceptibility testing was done using a BACTEC™ MGIT™ 960 Culture system (Becton Dickinson, USA) by standard protocol. Standard MTB genotyping methods, including spoligotyping and 24 loci MIRU-VNTR were applied to these strains as previously described [16] , [36] (link) (Table S2 ). Of them 28, 8, and 4 isolates had Beijing, LAM, and Ural spoligotype profiles, respectively. For Beijing isolates with spoligotype SIT1 IS6110 RFLP analysis was additionally performed [37] (link). BioNumerics version 5.1 package (Applied Maths, Belgium) was used for band comparison. According to RFLP analysis eight isolates belonged to Beijing B0/W148 cluster. Four of them were selected for a current whole genome re-sequencing project (Table 1 ).
3
Mycobacterium Culture and Drug Sensitivity
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The BACTEC MGIT 960 culture system (Becton, Dickinson and Company, America) was used for Mycobacterium culture. For positive results, strain identification was undertaken by Mpb64 (Hangzhou Genesis Biodetecton & Biocontrol Ltd, China) monoclonal antibody. First-line drug sensitivity was tested with BACTEC MGIT 960 system, and second-line drug sensitivity was tested with the absolute concentration method. Drug concentrations for isoniazid, rifampin, levofloxacin, moxifloxacin, amikacin, Capreomycin were 0.1, 1.0, 2.0, 1.0, 30, 40 μg/mL.11 The Ziehl-Neelsen acid-fast staining was used for the smear test.
4
Tuberculosis Nucleic Acid Detection Kit Protocol
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The M. tuberculosis Nucleic Acid Detection Kit (TB-SAT) (Nos. 20180801, 20181101, 20190301, 20190501, and 20190701), ABI 7500 real-time PCR instrument, and FZP-1 Nucleic Acid Purification Apparatus were ordered from Shanghai Rendu Biotechnology Co., Ltd. The BACTEC MGIT 960 culture system was obtained from Becton, Dickinson and Company (USA). A centrifugal machine was obtained from Hangzhou Xiangrui Technology Co., Ltd. A vortex mixer was purchased from HLD (Taicang, Shanghai, China). A dry bath incubator was ordered from Hangzhou Allsheng Instruments Co., Ltd.
5
Comprehensive Tuberculosis Diagnosis Protocol
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In clinical settings, all patients in possible TPE groups and MPE groups underwent TB-related tests. Samples used for M. tuberculosis culture were obtained from pleural fluid/sputum/lavage. The AFB smears were performed with auramine-rhodamine fluorochrome. The mycobacterial cultures were processed using the standard N-acetyl-L-cysteine and sodium hydroxide method using the BACTEC MGIT 960 culture system (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). PCR was performed using an M. tuberculosis kit (Sansur Biotech, China) for the detection of M. tuberculosis-complex on pleural biopsy and effusion samples. The PPD test was performed using 5 IU of tuberculin pure protein derivative (Xiangrui Biological Products Co. Ltd, China). Serum TB-antibody was measured using an immunochromatographic test system (Alere Inc., China). The ADA level of pleural fluid was evaluated using an ADA assay kit (Maccura Biotech, China). The IGRA assay was performed using a commercial kit from Oxford Immunotec Ltd (UK). Medical thoracoscopy was performed using an EVIS EXERA LTF-160 pleura videoscope manufactured by Olympus (Tokyo, Japan). Around four to six pieces were collected in formalin for histopathological examination. All tests were performed following the respective manufacturer’s instructions.
6
Tuberculosis Strain Characterization
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The RUS_B0 (earlier name 1947) (Beijing B0/W148 cluster) and the reference H37Rv strains of Mycobacterium tuberculosis were used. For proteomic and transcriptomic studies, M. tuberculosis strains were grown without shaking in liquid Middlebrook 7H9 medium with OADC supplement (Becton Dickinson, USA) at 37 °C in three biological replicates. To determine the growth rate, strains in 9 biological replicates were cultured in Mycobacterial growth indicator tubes (MGIT) on a BACTEC MGIT automated Mycobacterial detection system (Becton Dickinson) at 37 °C for 6–7 days. The susceptibility testing was done using the BACTEC MGIT 960 Culture system (Becton Dickinson) following the manufacturer’s protocol. Manipulation of cultures were performed in a BSC II (Thermo Fischer Scientific Inc., USA) within a TB-containment laboratory.
Whole-genome sequencing (WGS) data of 5,715 M. tuberculosis isolates was obtained from the National Center for Biotechnology Information (NCBI) and European Nucleotide Archive and used for B0 samples identification as reported previously11 (link).
Whole-genome sequencing (WGS) data of 5,715 M. tuberculosis isolates was obtained from the National Center for Biotechnology Information (NCBI) and European Nucleotide Archive and used for B0 samples identification as reported previously11 (link).
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