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Techniques for Isolation of Pure Cultures

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Microorganisms are routinely cultured in the laboratory using various techniques to isolate, grow, and quantify them for further study. These methods rely on inoculating microorganisms into a suitable growth medium under aseptic conditions to prevent contamination. Depending on the objective, inoculation can involve direct transfer or the use of diluted bacterial suspensions as the inoculum.Streak-Plate Method for IsolationThe streak-plate method is a common technique for obtaining pure...

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Updated: Oct 29, 2025

Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method

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Determining Minimum Inhibitory Concentrations in Liquid Cultures or on Solid Medium.

Qinglan Wang¹, Helena I M Boshoff²

¹Tuberculosis Research Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)

|July 8, 2021

Summary

This summary is machine-generated.

This chapter details methods for determining antimicrobial susceptibility of Mycobacterium tuberculosis. It covers testing in liquid and agar media under various conditions to aid tuberculosis drug development.

Keywords:

Acidified medium Agar MIC Carbon source Detergent Liquid MIC Minimum inhibitory concentration (MIC)Nitrosative stress

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Area of Science:

Microbiology
Pharmacology
Drug Discovery

Background:

Antimicrobial susceptibility testing is crucial for developing new tuberculosis drugs.
Standardized methods are needed to evaluate drug efficacy against Mycobacterium tuberculosis.

Purpose of the Study:

To describe various methods for determining the minimum inhibitory concentration (MIC) of compounds against Mycobacterium tuberculosis.
To outline susceptibility testing under diverse environmental conditions and media compositions.

Main Methods:

Determination of MIC in liquid media with varying carbon sources, detergents, and environmental stresses (acidic pH, reactive nitrogen intermediates).
Assessment of bovine serum albumin's effect on antimicrobial susceptibility in growth media.
Description of antimicrobial susceptibility testing on agar medium.

Main Results:

Established protocols for MIC determination against Mycobacterium tuberculosis.
Evaluated the impact of different media components and environmental factors on drug susceptibility.
Provided a comprehensive approach to antimicrobial susceptibility testing for tuberculosis drug development.

Conclusions:

The described methods provide a robust framework for evaluating novel antitubercular compounds.
Understanding susceptibility under varied conditions is essential for effective tuberculosis drug development programs.
These methods facilitate the screening and optimization of potential tuberculosis therapeutics.