Testing physiologically relevant conditions in minimal inhibitory concentration assays

Robert E. W. Hancock, Corrie R. Belanger

Published: 2021-07-01 DOI: 10.1038/s41596-021-00572-8

MIC assay

physiologically relevant conditions

antimicrobial susceptibility

broth microdilution

ESKAPE pathogens

AI 解读

Fabrication of angstrom-scale two-dimensional channels for mass transport

Efficient and strand-specific profiling of replicating chromatin with enrichment and sequencing of protein-associated nascent DNA in mammalian cells

The eINTACT method for studying nuclear changes in host plant cells targeted by bacterial effectors in native infection contexts

Rapid reaction optimization by robust and economical quantitative benchtop19F NMR spectroscopy

Profiling native pulmonary basement membrane stiffness using atomic force microscopy

Photoimmunotechnology as a powerful biological tool for molecular-based elimination of target cells and microbes, including bacteria, fungi and viruses

Measuring key human carbohydrate digestive enzyme activities using high-performance anion-exchange chromatography with pulsed amperometric detection

Neural cell isolation from adult macaques for high-throughput analyses and neurosphere cultures

Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds

Neuronal subtype-specific growth cone and soma purification from mammalian CNS via fractionation and fluorescent sorting for subcellular analyses and spatial mapping of local transcriptomes and proteomes

查看全部

Sections

Figures

References

The minimal inhibitory concentration (MIC) assay uses agar or broth dilution methods to measure, under defined test conditions, the lowest effective concentration of an antimicrobial agent that inhibits visible growth of a bacterium of interest. This assa
Introduction
Materials
Procedure
Troubleshooting
Timing
Anticipated results
References
ACKNOWLEDGMENTS

Wiegand, I., Hilpert, K. & Hancock, R. E. W. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat. Protoc. 3, 163–175 (2008).
Clinical Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing (CLSI document M100, 2020).
Nizet, V. The accidental orthodoxy of Drs. Mueller and Hinton. EBioMedicine 22, 26–27 (2017).
Cornforth, D. M., Diggle, F. L., Melvin, J. A., Bomberger, J. M. & Whiteley, M. Quantitative framework for model evaluation in microbiology research using Pseudomonas aeruginosa and cystic fibrosis infection as a test case. mBio 11, e03042-19 (2020).
Tata, M. et al. RNASeq based transcriptional profiling of Pseudomonas aeruginosa PA14 after short- and long-term anoxic cultivation in synthetic cystic fibrosis sputum medium. PLoS One 11, e0147811 (2016).
Palmer, K. L., Mashburn, L. M., Singh, P. K. & Whiteley, M. Cystic fibrosis sputum supports growth and cues key aspects of Pseudomonas aeruginosa physiology. J. Bacteriol 187, 5267–5277 (2005).
Kruczek, C. et al. Major transcriptome changes accompany the growth of Pseudomonas aeruginosa in blood from patients with severe thermal injuries. PLoS One 11, e0149229 (2016).
Umland, T. C. et al. In vivo-validated essential genes identified in Acinetobacter baumannii by using human ascites overlap poorly with essential genes detected on laboratory media. mBio 3, e00113-12 (2012).
Thulin, E., Thulin, M. & Andersson, D. I. Reversion of high-level mecillinam resistance to susceptibility in Escherichia coli during growth in urine. EBioMedicine 23, 111–118 (2017).
Lee, S. A. et al. General and condition-specific essential functions of Pseudomonas aeruginosa. Proc. Natl Acad. Sci. USA 112, 5189–5194 (2015).
Ersoy, S. C. et al. Correcting a fundamental flaw in the paradigm for antimicrobial susceptibility testing. EBioMedicine 20, 173–181 (2017).
Hancock, R. E. W. Rethinking the antibiotic discovery paradigm. EBioMedicine 2, 629–630 (2015).
Kubicek-Sutherland, J. Z. et al. Host-dependent induction of transient antibiotic resistance: a prelude to treatment failure. EBioMedicine 2, 1169–1178 (2015).
World Health Organization. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. http://www.who.int/medicines/publications/global-priority-list-antibiotic-resistant-bacteria/en/ (WHO, 2017).
De Oliveira, D. M. P. et al. Antimicrobial resistance in ESKAPE pathogens. Clin. Microbiol. Rev. 33, e00181-19 (2020).
Turner, K. H., Wessel, A. K., Palmer, G. C., Murray, J. L. & Whiteley, M. Essential genome of Pseudomonas aeruginosa in cystic fibrosis sputum. Proc. Natl Acad. Sci. 112, 4110–4115 (2015).
Malachowa, N. et al. Global changes in Staphylococcus aureus gene expression in human blood. PLoS One 6, e18617 (2011).
Quinn, B. et al. Human serum albumin alters specific genes that can play a role in survival and persistence in Acinetobacter baumannii. Sci. Rep. 8, 14741 (2018).
Mäder, U. et al. Staphylococcus aureus transcriptome architecture: from laboratory to infection-mimicking conditions. PLoS Genet 12, e1005962 (2016).
Belanger, C. R. et al. Identification of novel targets of azithromycin activity against Pseudomonas aeruginosa grown in physiologically relevant media. Proc. Natl Acad. Sci. 117, 33519–33529 (2020).
Winsor, G. L. et al. Enhanced annotations and features for comparing thousands of Pseudomonas genomes in the Pseudomonas genome database. Nucleic Acids Res. 44, D646–D653 (2016).
Fung, C. et al. Gene expression of Pseudomonas aeruginosa in a mucin-containing synthetic growth medium mimicking cystic fibrosis lung sputum. J. Med. Microbiol. 59, 1089–1100 (2010).
Lin, L. et al. Azithromycin synergizes with cationic antimicrobial peptides to exert bactericidal and therapeutic activity against highly multidrug-resistant Gram-negative bacterial pathogens. EBioMedicine 2, 690–698 (2015).
Colquhoun, J. M., Wozniak, R. A. F. & Dunman, P. M. Clinically relevant growth conditions alter Acinetobacter baumannii antibiotic susceptibility and promote identification of novel antibacterial agents. PLoS One 10, e0143033 (2015).
Yeaman, M. R., Gank, K. D., Bayer, A. S. & Brass, E. P. Synthetic peptides that exert antimicrobial activities in whole blood and blood-derived matrices. Antimicrob. Agents Chemother. 46, 3883–3891 (2002).
Weber, B. S. et al. Genetic and chemical screening in human blood serum reveals unique antibacterial targets and compounds against Klebsiella pneumoniae. Cell Rep 32, 107927 (2020).
Palmer, K. L., Aye, L. M. & Whiteley, M. Nutritional cues control Pseudomonas aeruginosa multicellular behavior in cystic fibrosis sputum. J. Bacteriol. 189, 8079–8087 (2007).
Kirchner, S. et al. Use of artificial sputum medium to test antibiotic efficacy against Pseudomonas aeruginosa in conditions more relevant to the cystic fibrosis lung. J. Vis. Exp. https://doi.org/10.3791/3857 (2012).
Sriramulu, D. D., Lünsdorf, H., Lam, J. S. & Römling, U. Microcolony formation: a novel biofilm model of Pseudomonas aeruginosa for the cystic fibrosis lung. J. Med. Microbiol. 54, 667–676 (2005).
Yeung, A. T. Y., Parayno, A. & Hancock, R. E. W. Mucin promotes rapid surface motility in Pseudomonas aeruginosa. mBio 3, e00073–12 (2012).
Kreda, S. M., Davis, C. W. & Rose, M. C. CFTR, mucins, and mucus obstruction in cystic fibrosis. Cold Spring Harb. Perspect. Med. 2, a009589 (2012).
Kirchner, K. K., Wagener, J. S., Khan, T. Z., Copenhaver, S. C. & Accurso, F. J. Increased DNA levels in bronchoalveolar lavage fluid obtained from infants with cystic fibrosis. Am. J. Respir. Crit. Care Med. 154, 1426–1429 (1996).
Haney, E., Trimble, M., Cheng, J., Vallé, Q. & Hancock, R. Critical assessment of methods to quantify biofilm growth and evaluate antibiofilm activity of host defence peptides. Biomolecules 8, 29 (2018).

Testing physiologically relevant conditions in minimal inhibitory concentration assays

Supplementary information

推荐阅读