Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry

Kieran Mulroney; Margaret Kopczyk; Christine Carson; Teagan Paton; Timothy Inglis; Aron Chakera

doi:10.1016/j.ebiom.2022.104145

Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry

Kieran Mulroney, Margaret Kopczyk, Christine Carson, Teagan Paton, Timothy Inglis, Aron Chakera

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

Abstract

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.

Original language	English
Article number	104145
Journal	EBioMedicine
Volume	82
DOIs	https://doi.org/10.1016/j.ebiom.2022.104145
Publication status	Published - Aug 2022

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10.1016/j.ebiom.2022.104145Licence: CC BY-NC-ND

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@article{decf64382ad9440d9273415aeb674aa0,

title = "Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry",

abstract = "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.",

keywords = "Antimicrobial susceptibility, Flow cytometry, PD, Peritoneal dialysis, Peritonitis, Rapid diagnostics",

author = "Kieran Mulroney and Margaret Kopczyk and Christine Carson and Teagan Paton and Timothy Inglis and Aron Chakera",

note = "Funding Information: The authors gratefully acknowledge the provision of clinical specimens and microbiological strains by PathWest Laboratory Medicine WA. Hardware and consumable support were provided by Dr Michael Ward, of Life Technologies (ThermoFisher Scientific). The authors thank Professor Nicholas Topley for scientific and technical review of the project and manuscript. The work received funding from the Government of Western Australia, Department of Health via a Research Translation Project Grant (RTP2019R13), from the Government of Australia, National Health and Medical Research Council Ideas Grant (APP 2004298), and the Forrest Research Foundation via a Forrest Prospect Fellowship Awarded to Dr Kieran Mulroney. Funding Information: The authors gratefully acknowledge the provision of clinical specimens and microbiological strains by PathWest Laboratory Medicine WA. Hardware and consumable support were provided by Dr Michael Ward, of Life Technologies (ThermoFisher Scientific). The authors thank Professor Nicholas Topley for scientific and technical review of the project and manuscript. The work received funding from the Government of Western Australia, Department of Health via a Research Translation Project Grant (RTP2019R13), from the Government of Australia, National Health and Medical Research Council Ideas Grant (APP 2004298), and the Forrest Research Foundation via a Forrest Prospect Fellowship Awarded to Dr Kieran Mulroney. Publisher Copyright: {\textcopyright} 2022 The Authors",

year = "2022",

month = aug,

doi = "10.1016/j.ebiom.2022.104145",

language = "English",

volume = "82",

journal = "EBioMedicine",

issn = "2352-3964",

publisher = "Elsevier",

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T1 - Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry

AU - Mulroney, Kieran

AU - Kopczyk, Margaret

AU - Carson, Christine

AU - Paton, Teagan

AU - Inglis, Timothy

AU - Chakera, Aron

N1 - Funding Information: The authors gratefully acknowledge the provision of clinical specimens and microbiological strains by PathWest Laboratory Medicine WA. Hardware and consumable support were provided by Dr Michael Ward, of Life Technologies (ThermoFisher Scientific). The authors thank Professor Nicholas Topley for scientific and technical review of the project and manuscript. The work received funding from the Government of Western Australia, Department of Health via a Research Translation Project Grant (RTP2019R13), from the Government of Australia, National Health and Medical Research Council Ideas Grant (APP 2004298), and the Forrest Research Foundation via a Forrest Prospect Fellowship Awarded to Dr Kieran Mulroney. Funding Information: The authors gratefully acknowledge the provision of clinical specimens and microbiological strains by PathWest Laboratory Medicine WA. Hardware and consumable support were provided by Dr Michael Ward, of Life Technologies (ThermoFisher Scientific). The authors thank Professor Nicholas Topley for scientific and technical review of the project and manuscript. The work received funding from the Government of Western Australia, Department of Health via a Research Translation Project Grant (RTP2019R13), from the Government of Australia, National Health and Medical Research Council Ideas Grant (APP 2004298), and the Forrest Research Foundation via a Forrest Prospect Fellowship Awarded to Dr Kieran Mulroney. Publisher Copyright: © 2022 The Authors

PY - 2022/8

Y1 - 2022/8

N2 - 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.

AB - 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.

KW - Antimicrobial susceptibility

KW - Flow cytometry

KW - PD

KW - Peritoneal dialysis

KW - Peritonitis

KW - Rapid diagnostics

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DO - 10.1016/j.ebiom.2022.104145

M3 - Article

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AN - SCOPUS:85135453712

SN - 2352-3964

VL - 82

JO - EBioMedicine

JF - EBioMedicine

M1 - 104145

ER -

Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry

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Culture-independent microbiology: reducing delays in the diagnosis of severe infections and detection of antimicrobial resistance from days to hours

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Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry

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Culture-independent microbiology: reducing delays in the diagnosis of severe infections and detection of antimicrobial resistance from days to hours

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