Chapter 28
Antimicrobial Susceptibility Testing for Mycobacterium sps
in High-Throughput Format
Esther Pe ´ rez-Herra ´ n and Alfonso Mendoza
Abstract
The concept of antimicrobial susceptibility testing is an essential part of clinical microbiology. Antimicrobial
testing has played a central role in the identification of new antibiotics and defining their clinical uses. Here
we describe different approaches to determine the activity of compounds in medium or high-throughput
format.
Key words Antimicrobial susceptibility test, HTS whole cell assay, Minimum inhibitory concentration
(MIC), Resazurin, ATP measurement
1 Introduction
Tuberculosis (TB) is one of the leading causes of death in the world.
In addition, multidrug-resistant (MDR)-TB incidence is rising in
many areas around the world with 5–10% being extensively drugresistant (XDR). The development of new types of antitubercular
drugs is urgently needed [1]. Nowadays, most, if not all of the
known antituberculars have been identified through phenotypic
assays, and the development of fast and reliable ways to determine
the inhibitory activity of chemical entities is still a must. The methods described below have been adapted to screen the activity of
thousands of compounds against Mycobacterium tuberculosis under
replicating conditions [2] and intracellularly [3].
At the dawn of microbiology, the identification of substances
that induced or blocked the growth of microorganisms led to the
development of methods for the quantitative estimation of these
growth-promoting or growth-inhibiting substances. Many references to antibiosis can be found in the works of Pasteur, Koch,
and Paul Ehrlich. Their observations were supported by experimental evidence (from in vitro growth inhibition to animal
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_28, © Springer Science+Business Media, LLC, part of Springer Nature 2021
637
Antimicrobial Susceptibility Testing for Mycobacterium sps
in High-Throughput Format
Esther Pe ´ rez-Herra ´ n and Alfonso Mendoza
Abstract
The concept of antimicrobial susceptibility testing is an essential part of clinical microbiology. Antimicrobial
testing has played a central role in the identification of new antibiotics and defining their clinical uses. Here
we describe different approaches to determine the activity of compounds in medium or high-throughput
format.
Key words Antimicrobial susceptibility test, HTS whole cell assay, Minimum inhibitory concentration
(MIC), Resazurin, ATP measurement
1 Introduction
Tuberculosis (TB) is one of the leading causes of death in the world.
In addition, multidrug-resistant (MDR)-TB incidence is rising in
many areas around the world with 5–10% being extensively drugresistant (XDR). The development of new types of antitubercular
drugs is urgently needed [1]. Nowadays, most, if not all of the
known antituberculars have been identified through phenotypic
assays, and the development of fast and reliable ways to determine
the inhibitory activity of chemical entities is still a must. The methods described below have been adapted to screen the activity of
thousands of compounds against Mycobacterium tuberculosis under
replicating conditions [2] and intracellularly [3].
At the dawn of microbiology, the identification of substances
that induced or blocked the growth of microorganisms led to the
development of methods for the quantitative estimation of these
growth-promoting or growth-inhibiting substances. Many references to antibiosis can be found in the works of Pasteur, Koch,
and Paul Ehrlich. Their observations were supported by experimental evidence (from in vitro growth inhibition to animal
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_28, © Springer Science+Business Media, LLC, part of Springer Nature 2021
637