FAST: Rapid determinations of antibiotic susceptibility phenotypes using label-free cytometry

Cytometry A. 2018 Jun;93(6):639-648. doi: 10.1002/cyto.a.23370. Epub 2018 May 7.

Abstract

Sepsis, a life-threatening immune response to blood infections (bacteremia), has a ∼30% mortality rate and is the 10th leading cause of US hospital deaths. The typical bacterial loads in adult septic patients are ≤100 bacterial cells (colony forming units, CFU) per ml blood, while pediatric patients exhibit only ∼1000 CFU/ml. Due to the low numbers, bacteria must be propagated through ∼24-hours blood cultures to generate sufficient CFUs for diagnosis and further analyses. Herein, we demonstrate that, unlike other rapid post-blood culture antibiotic susceptibility tests (ASTs), our phenotypic approach can drastically accelerate ASTs for the most common sepsis-causing gram-negative pathogens by circumventing long blood culture-based amplification. For all blood isolates of multi-drug resistant pathogens investigated (Escherichia coli, Klebsiella pneumoniae, and Acinetobacter nosocomialis), effective antibiotic(s) were readily identified within the equivalent of 8 hours from initial blood draw using <0.5 mL of adult blood per antibiotic. These methods should drastically improve patient outcomes by significantly reducing time to actionable treatment information and reduce the incidence of antibiotic resistance. © 2018 International Society for Advancement of Cytometry.

Keywords: antibiotic susceptibility test; flow cytometry; multidimensional statistics; multidrug-resistant bacteria; pre-blood culture.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacteremia / blood
  • Bacteremia / drug therapy
  • Cells, Cultured
  • Drug Resistance, Multiple, Bacterial / physiology*
  • Erythrocytes / drug effects
  • Erythrocytes / microbiology*
  • Erythrocytes / physiology*
  • Escherichia coli / drug effects
  • Escherichia coli / isolation & purification
  • Flow Cytometry / methods*
  • Humans
  • Microbial Sensitivity Tests
  • Phenotype*
  • Time Factors

Substances

  • Anti-Bacterial Agents