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You will know by its tail: a method for quantification of heterogeneity of bacterial populations using single cell MIC profiling

Natalia Pacocha, Marta Zapotoczna, Karol Makuch, Jakub Bogusławski, Piotr Garstecki
doi: https://doi.org/10.1101/2022.04.29.490018
Natalia Pacocha
1Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland
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Marta Zapotoczna
2Department of Molecular Microbiology, Institute of Microbiology, Faculty of Biology, Biological and Chemical Research Centre, University of Warsaw, Żwirki i Wigury 101, 02-089 Warsaw, Poland
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Karol Makuch
1Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland
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  • For correspondence: kmakuch@ichf.edu.pl garst@ichf.edu.pl
Jakub Bogusławski
3International Centre for Translational Eye Research, Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland
4Department of Physical Chemistry of Biological Systems, Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52,01-224 Warsaw, Poland
5Laser & Fiber Electronics Group, Faculty of Electronics, Wrocław University of Science and Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland
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Piotr Garstecki
1Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland
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  • For correspondence: kmakuch@ichf.edu.pl garst@ichf.edu.pl
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Abstract

Severe non-healing infections are often caused by multiple pathogens or by genetic variants of the same pathogen exhibiting different levels of antibiotic resistance. For example, polymicrobial diabetic foot infections double the risk of amputation compared to monomicrobial infections. Although these infections lead to increased morbidity and mortality, standard antimicrobial susceptibility methods are designed for homogenous samples and are impaired in quantifying heteroresistance. Here, we propose a droplet-based label-free method for quantifying the antibiotic response of the entire population at the single-cell level. We used Pseudomonas aeruginosa and Staphylococcus aureus samples to confirm that the shape of the profile informs about the coexistence of diverse bacterial subpopulations, their sizes, and antibiotic heteroresistance. These profiles could therefore indicate the outcome of antibiotic treatment in terms of the size of remaining subpopulations. Moreover, we studied phenotypic variants of a S. aureus strain to confirm that the profile can be used to identify tolerant subpopulations, such as small colony variants, associated with increased risks for the development of persisting infections. Therefore, the profile is a versatile instrument for quantifying the size of each bacterial subpopulation within a specimen as well as their individual and joined heteroresistance.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 29, 2022.
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You will know by its tail: a method for quantification of heterogeneity of bacterial populations using single cell MIC profiling
Natalia Pacocha, Marta Zapotoczna, Karol Makuch, Jakub Bogusławski, Piotr Garstecki
bioRxiv 2022.04.29.490018; doi: https://doi.org/10.1101/2022.04.29.490018
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You will know by its tail: a method for quantification of heterogeneity of bacterial populations using single cell MIC profiling
Natalia Pacocha, Marta Zapotoczna, Karol Makuch, Jakub Bogusławski, Piotr Garstecki
bioRxiv 2022.04.29.490018; doi: https://doi.org/10.1101/2022.04.29.490018

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