MIC assays were conducted using Sensititre MYCOTB MIC plates (Thermo Fisher Scientific) according to the manufacturer’s instructions. In brief, the MTB isolate was first subcultured on 7H10 Agar (Becton, Dickinson and Co., Sparks, MD). Multiple colonies were resuspended with glass beads in a saline-Tween solution, and the turbidity was adjusted to a McFarland standard of 0.5. A 100-μL volume of the resuspension was next mixed with 11 mL 7H9 broth containing oleic acid albumin-dextrose-catalase (Trek Diagnostic Systems, Cleveland, OH). A 100-μL volume of diluent was inoculated in each well of the MYCOTB plate and sealed with a permanent plastic seal at 37°C in 5% CO2. Plates were monitored on days 7, 10, 14, and 21 using a mirrored viewer. The lowest concentration with no visible growth was considered the MIC of rifampin or rifabutin.
Oleic acid albumin dextrose catalase
Manufactured by Thermo Fisher Scientific
Sourced in United States
Oleic acid albumin-dextrose-catalase is a culture media supplement used in microbiology and cell culture applications. It provides nutrients and growth factors to support the growth and maintenance of various cell types and microbial cultures.
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5 protocols using oleic acid albumin dextrose catalase
1
Mycobacterium tuberculosis MIC Assay
2
Mycobacterium tuberculosis MIC Assay
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MIC assays were conducted using Sensititre MYCOTB MIC plates (Thermo Fisher Scientific) according to the manufacturer’s instructions. In brief, the MTB isolate was first subcultured on 7H10 Agar (Becton, Dickinson and Co., Sparks, MD). Multiple colonies were resuspended with glass beads in a saline-Tween solution, and the turbidity was adjusted to a McFarland standard of 0.5. A 100-μL volume of the resuspension was next mixed with 11 mL 7H9 broth containing oleic acid albumin-dextrose-catalase (Trek Diagnostic Systems, Cleveland, OH). A 100-μL volume of diluent was inoculated in each well of the MYCOTB plate and sealed with a permanent plastic seal at 37°C in 5% CO2. Plates were monitored on days 7, 10, 14, and 21 using a mirrored viewer. The lowest concentration with no visible growth was considered the MIC of rifampin or rifabutin.
3
Mycobacterium Tuberculosis Antibiotic Susceptibility
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Tests were conducted using Sensititre MYCOTB MIC plates (ThermoFisher Scientific) according to the manufacturer’s instructions. In brief, an MTB isolate was subcultured on 7H10 agar (Becton, Dickinson and Co., Sparks, MD, USA). Fresh colonies were resuspended with glass beads in a saline-Tween solution, and the turbidity was adjusted to a McFarland standard of 0.5. After 15 min, 100 µl of the resuspension was poured and mixed with 11 ml 7H9 broth containing oleic acid albumin-dextrose-catalase (Trek Diagnostic Systems, Cleveland, OH). Diluent (100 µl) was inoculated into each well of the MYCOTB plate that was covered with permanent plastic seals and incubated at 37 °C in 5% CO2. Plates were monitored on days 7, 10, 14, and 21 using a mirrored viewer. No visible growth with the lowest concentration was considered the MIC of each antibiotic.
4
Isolation and Identification of MABC
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Between 2014 and 2018, 129 MABC isolates were randomly isolated from the sputum and bronchoalveolar fluids of patients with clinical signs of lower respiratory tract infections in three tertiary hospitals (Shanghai Pulmonary Hospital, the Second Affiliated Hospital of Nanchang University, and the Second Affiliated Hospital of Suzhou University). All isolates were cultured in Middlebrook 7H10broth (BD, France) supplemented with 10% (vol/vol) oleic acid-albumin-dextrose-catalase (Thermo Fisher Scientific, USA). The cultures were incubated at 37°C for 7 days.
The Ethics Committee of Shanghai Pulmonary Hospital, Tongji University School of Medicine, exempted this study from ethical review because the assessment of the bacteria was part of routine hospital laboratory procedures. Verbal informed consent was obtained from all participants.
The Ethics Committee of Shanghai Pulmonary Hospital, Tongji University School of Medicine, exempted this study from ethical review because the assessment of the bacteria was part of routine hospital laboratory procedures. Verbal informed consent was obtained from all participants.
5
Culturing Mycobacterium tuberculosis Erdman
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