A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections
Journal
Biosensors
Series/Report No.
Biosensors
Journal Volume
11
Journal Issue
8
Start Page
288
ISSN
2079-6374
Date Issued
2021-08-22
Author(s)
Forsyth, Britney
Torab, Peter
Malcom, Tyler
Wang, Tza-Huei
Liao, Joseph C.
Yang, Samuel
Kvam, Erik
Puleo, Chris
Wong, Pak Kin
Abstract
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.
Subjects
Diagnostics
Microfluidics
Multidrug-resistant bacteria
Sepsis
Single-cell analysis
Publisher
MDPI AG
Type
journal article