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…

Offenlegungen

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

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Evaluating the Effectiveness of a Test Compound against Mycobacteria in Broth. J. Vis. Exp. (Pending Publication), e21978, doi: (2024).

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