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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Bibliographic Details
Published in:Biosensors (Basel) 2021-08, Vol.11 (8), p.288
Main Authors: Forsyth, Britney, Torab, Peter, Lee, Jyong-Huei, Malcom, Tyler, Wang, Tza-Huei, Liao, Joseph C, Yang, Samuel, Kvam, Erik, Puleo, Chris, Wong, Pak Kin
Format: Article
Language:English
Subjects:
Anti-Bacterial Agents
Antibiotic resistance
Antibiotics
Antiinfectives and antibacterials
Antimicrobial agents
Automation
Bacteria
Bacterial diseases
Bacterial Infections
Blood
Blood levels
Cell culture
Centrifugation
Depletion
Dextran
Dextrans
diagnostics
E coli
Efficiency
Enrichment
Erythrocytes
Humans
Infections
Lab-On-A-Chip Devices
Laboratories
Microbial Sensitivity Tests
Microfluidics
Morbidity
multidrug-resistant bacteria
Pathogens
Patients
Plasma
Polymerase chain reaction
Sepsis
Sepsis - diagnosis
Silicon wafers
single-cell analysis
Workflow
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Summary: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.
ISSN:2079-6374
2079-6374
DOI:10.3390/bios11080288