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Updated: Sep 4, 2025
One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures
Published on: July 9, 2012
Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry
Kieran Mulroney1, Margaret Kopczyk2, Christine Carson3
1Harry Perkins Institute of Medical Research, 6 Verdun Street, Nedlands, WA 6009, Australia; School of Medicine, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
Background:
The annual mortality burden of antimicrobial resistant infections exceeds 1.27 million/year. With serious infections, every hour without effective antimicrobial therapy results in a 6.7% increased risk of death. New technology that delivers actionable pathology results in clinically-relevant timeframes is an urgent priority. We present the development and validation of an acoustic-enhanced flow cytometric (AFC) workflow that provides same-day confirmation of infection and antimicrobial susceptibility, using peritoneal dialysis (PD)-associated peritonitis as a demonstrative example.
Methods:
In this cohort study, we analysed peritoneal dialysis effluent specimens using AFC to confirm the presence of infection and antimicrobial susceptibility of identified organisms. The primary outcome was the performance of the assay compared to conventional microbiology performed by the clinical laboratory. A secondary outcome was time to result.
Findings:
AFC confirmed infection from primary specimens (n=116), with a sensitivity of 86% and specificity of 94% in ≤ one hour from arrival, including confirmation of infecting organisms in culture-negative cases. Combined with flow-cytometry-assisted antimicrobial susceptibility testing (FAST), we demonstrate same-day antimicrobial susceptibility profiles with an accuracy equivalent to conventional laboratory-based tests.
Interpretation:
Application of AFC based assays to confirm infection and predict antimicrobial susceptibility can deliver actionable results with a performance that meets or exceeds currently utilised microbiological tests in clinically meaningful timeframes, as demonstrated for PD peritonitis. This technology shows potential for broad applicability to other time-critical serious infections.
Funding:
Government of Western Australia (Department of Health), Government of Australia (National Health and Medical Research Council) and the Forrest Research Foundation.
Insights
A new acoustic-enhanced flow cytometry (AFC) method rapidly confirms infections and antimicrobial susceptibility within hours. This technology offers same-day results, improving patient outcomes for serious infections like peritoneal dialysis peritonitis.
Area of Science:
- Microbiology
- Flow Cytometry
- Medical Diagnostics
Background:
- Antimicrobial resistance causes over 1.27 million deaths annually.
- Delayed diagnosis and treatment of serious infections increase mortality risk.
- Rapid diagnostic technologies are crucial for effective antimicrobial therapy.
Purpose of the Study:
- To develop and validate an acoustic-enhanced flow cytometry (AFC) workflow.
- To provide same-day confirmation of infection and antimicrobial susceptibility.
- To demonstrate the workflow's utility using peritoneal dialysis (PD)-associated peritonitis.
Main Methods:
- A cohort study analyzed peritoneal dialysis effluent specimens using AFC.
- AFC was used to confirm infection and determine antimicrobial susceptibility.
- Assay performance was compared to conventional microbiology, with time-to-result as a secondary outcome.
Main Results:
- AFC confirmed infection in 116 primary specimens with 86% sensitivity and 94% specificity within one hour.
- Infecting organisms were identified even in culture-negative cases.
- Combined with flow-cytometry-assisted antimicrobial susceptibility testing (FAST), same-day susceptibility profiles were achieved with accuracy comparable to conventional methods.
Conclusions:
- AFC-based assays deliver actionable results for infection confirmation and antimicrobial susceptibility prediction.
- The technology meets or exceeds current microbiological test performance in clinically relevant timeframes.
- AFC shows potential for broad application in other time-critical serious infections.
