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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-8391654 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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