Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria

Ireneusz Rosłon; Aleksandre Japaridze; Stef Rodenhuis; Lieke Hamoen; Murali Krishna Ghatkesar; Peter Steeneken; Cees Dekker; Farbod Alijani

doi:10.1016/j.isci.2023.108268

Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria

Ireneusz Rosłon, Aleksandre Japaridze, Stef Rodenhuis, Lieke Hamoen, Murali Krishna Ghatkesar, Peter Steeneken, Cees Dekker, Farbod Alijani^*

^*Corresponding author for this work

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Abstract

Bacteria that are resistant to antibiotics present an increasing burden on healthcare. To address this emerging crisis, novel rapid antibiotic susceptibility testing (AST) methods are eagerly needed. Here, we present an optical AST technique that can determine the bacterial viability within 1 h down to a resolution of single bacteria. The method is based on measuring intensity fluctuations of a reflected laser focused on a bacterium in reflective microwells. Using numerical simulations, we show that both refraction and absorption of light by the bacterium contribute to the observed signal. By administering antibiotics that kill the bacteria, we show that the variance of the detected fluctuations vanishes within 1 h, indicating the potential of this technique for rapid sensing of bacterial antibiotic susceptibility. We envisage the use of this method for massively parallelizable AST tests and fast detection of drug-resistant pathogens.

Original language	English
Article number	108268
Number of pages	11
Journal	iScience
Volume	26
Issue number	11
DOIs	https://doi.org/10.1016/j.isci.2023.108268
Publication status	Published - 2023

Funding

We acknowledge Michiel Otte and Le-Vaughn Naarden for their help in performing measurements with deep cavities. We acknowledge Leo Smeets for providing the clinical isolates. Financial support was provided from the European Union's Horizon 2020 research and innovation programme under ERC starting grant ENIGMA (no. 802093, F.A. and I.E.R.), ERC PoC GRAPHFITI (no. 966720, F.A. and A.J.), Dutch Research Council (NWO) take-off grant (no. 19639), Graphene Flagship (grant nos. 785219 and 881603, P.S.), and the ERC Advanced Grant LoopingDNA (no. 883684, C.D.), as well as NWO/OCW as part of the NanoFront and BaSyC programmes and by the Swiss National Science Foundation (grant no. P300P2_177768, A.J.). I.E.R. A.J. and F.A. conceived the idea. I.E.R. A.J. and S.R. collected the data and performed the interferometry experiments. I.E.R. constructed the setup and performed the simulations. A.J. and L.H. performed the bacterial manipulation. All authors designed the experiments. The project was supervised by F.A. M.G. C.D. and P.G.S. All authors contributed to the data analysis, interpretation of the results, and writing of the manuscript. Employment or leadership: A.J.; SoundCell B.V. Consultant or advisory role: I.E.R. P.G.S. and F.A.; SoundCell B.V. The authors declare no further competing interests.

Keywords

Medical Microbiology
Microbiology

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10.1016/j.isci.2023.108268

PIIS2589004223023453Final published version, 3.16 MBLicence: CC BY

Cite this

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title = "Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria",

abstract = "Bacteria that are resistant to antibiotics present an increasing burden on healthcare. To address this emerging crisis, novel rapid antibiotic susceptibility testing (AST) methods are eagerly needed. Here, we present an optical AST technique that can determine the bacterial viability within 1 h down to a resolution of single bacteria. The method is based on measuring intensity fluctuations of a reflected laser focused on a bacterium in reflective microwells. Using numerical simulations, we show that both refraction and absorption of light by the bacterium contribute to the observed signal. By administering antibiotics that kill the bacteria, we show that the variance of the detected fluctuations vanishes within 1 h, indicating the potential of this technique for rapid sensing of bacterial antibiotic susceptibility. We envisage the use of this method for massively parallelizable AST tests and fast detection of drug-resistant pathogens.",

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AU - Rosłon, Ireneusz

AU - Japaridze, Aleksandre

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AU - Ghatkesar, Murali Krishna

AU - Steeneken, Peter

AU - Dekker, Cees

AU - Alijani, Farbod

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AB - Bacteria that are resistant to antibiotics present an increasing burden on healthcare. To address this emerging crisis, novel rapid antibiotic susceptibility testing (AST) methods are eagerly needed. Here, we present an optical AST technique that can determine the bacterial viability within 1 h down to a resolution of single bacteria. The method is based on measuring intensity fluctuations of a reflected laser focused on a bacterium in reflective microwells. Using numerical simulations, we show that both refraction and absorption of light by the bacterium contribute to the observed signal. By administering antibiotics that kill the bacteria, we show that the variance of the detected fluctuations vanishes within 1 h, indicating the potential of this technique for rapid sensing of bacterial antibiotic susceptibility. We envisage the use of this method for massively parallelizable AST tests and fast detection of drug-resistant pathogens.

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Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria

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Microwell-enhanced optical rapid antibiotic susceptibility testing of single bacteria

Abstract

Funding

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Dataset underlying the publication: Microwell-enhanced optical detection of single bacteria

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