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A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections

Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and th...

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Autores principales: Forsyth, Britney, Torab, Peter, Lee, Jyong-Huei, Malcom, Tyler, Wang, Tza-Huei, Liao, Joseph C., Yang, Samuel, Kvam, Erik, Puleo, Chris, Wong, Pak Kin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8391654/
https://www.ncbi.nlm.nih.gov/pubmed/34436090
http://dx.doi.org/10.3390/bios11080288
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author Forsyth, Britney
Torab, Peter
Lee, Jyong-Huei
Malcom, Tyler
Wang, Tza-Huei
Liao, Joseph C.
Yang, Samuel
Kvam, Erik
Puleo, Chris
Wong, Pak Kin
author_facet Forsyth, Britney
Torab, Peter
Lee, Jyong-Huei
Malcom, Tyler
Wang, Tza-Huei
Liao, Joseph C.
Yang, Samuel
Kvam, Erik
Puleo, Chris
Wong, Pak Kin
author_sort Forsyth, Britney
collection PubMed
description Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20–30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours.
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spelling pubmed-83916542021-08-28 A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections Forsyth, Britney Torab, Peter Lee, Jyong-Huei Malcom, Tyler Wang, Tza-Huei Liao, Joseph C. Yang, Samuel Kvam, Erik Puleo, Chris Wong, Pak Kin Biosensors (Basel) Article Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20–30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours. MDPI 2021-08-22 /pmc/articles/PMC8391654/ /pubmed/34436090 http://dx.doi.org/10.3390/bios11080288 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Forsyth, Britney
Torab, Peter
Lee, Jyong-Huei
Malcom, Tyler
Wang, Tza-Huei
Liao, Joseph C.
Yang, Samuel
Kvam, Erik
Puleo, Chris
Wong, Pak Kin
A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title_full A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title_fullStr A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title_full_unstemmed A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title_short A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
title_sort rapid single-cell antimicrobial susceptibility testing workflow for bloodstream infections
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8391654/
https://www.ncbi.nlm.nih.gov/pubmed/34436090
http://dx.doi.org/10.3390/bios11080288
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