Evaluating the Effectiveness of a Test Compound against Mycobacteria in Broth
Published: February 29, 2024
Abstract
Source: Zheng, X., et al. System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis. J. Vis. Exp. (2017)
This video demonstrates an in-broth assay to determine antibiotic efficacy against Mycobacterium tuberculosis. The bacterial culture is incubated with different concentrations of a bactericidal antibiotic. The bactericidal efficacy is measured by assessing the presence of viable bacteria using a resazurin assay.
Protocol
1. Bacterial Strain and Growth Medium Make albumin dextrose and salt stock solution (ADS) by solubilizing 25 g of bovine serum albumin, 10.0 g of dextrose, and 4.05 g of sodium chloride in 460 mL of deionized water. Filter-sterilize the ADS and store it at 4 °C. Make 7H9 broth by adding 4.7 g of 7H9 powder and 2 mL of glycerol to 900 mL of purified water. Autoclave the 7H9 broth at 121 °C for 10 min and allow it to cool to room temperature before proceeding. Make 7H9ADST b…
Divulgaciones
The authors have nothing to disclose.
Materials
| Middlebrook 7H9 | Becton, Dickinson and Company | 271210 | |
| Tween80 | Fisher Scientific | T164 | |
| Albumin, Bovine pH7 | Affymetrix | 10857 | |
| Dextrose | Fisher Scientific | BP350 | |
| Sodium Chloride | Fisher Scientific | BP358 | |
| kanamycin sulfate | Fisher Scientific | BP906 | |
| Resazurin | Alfa Aesar | B21187 | |
| Glycerol | Fisher Scientific | BP229 | |
| M. tuberculosis H37Rv | |||
| 96-well flat bottom white plate | Corning | 3917 | |
| 95-well flat bottom clear plate | Corning | 3595 | |
| Transparent plate sealer | Thermo Fisher Scientific | AB-0580 | |
| Luminometer | Applied Biosystems | Tropix TR717 |
Tags
Citar este artículo
Evaluating the Effectiveness of a Test Compound against Mycobacteria in Broth. J. Vis. Exp. (Pending Publication), e21978, doi: (2024).