Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin

Collette Allen; Carrie Turner; Sumit Kalsi; David Jamieson; Yuetao Li; Hywel Morgan; John Sutton

doi:10.1016/j.diagmicrobio.2019.114926

Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin

Collette Allen, Carrie Turner, Sumit Kalsi, David Jamieson, Yuetao Li, Hywel Morgan, John Sutton^*

^*Corresponding author for this work

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

1 Citation (Scopus)

Abstract

Routine identification of carbapenemase-producing bacterial isolates is a lengthy process often taking up to 72 h to generate results with standard culture-based tests. Here we describe a rapid test based on the hydrolysis of nitrocefin to identify isolates producing β-lactamase enzymes. A cocktail of inhibitors has been optimized in the reaction mix to provide specificity for carbapenemase enzymes. The developed assay has also been translated to a microfluidic platform with an optical readout (optofluidic chip). The chip has a long absorbance path (25 mm) to provide high sensitivity. A sample-to-answer has been achieved in under 30 min on these chips using colonies from culture plates. The test on this platform has the potential to provide a rapid indicative (presumptive positive) test for carbapenemase producers direct from bacteria isolated from patient samples, to rapidly trigger infection control measures and identify samples that should be prioritized for more specialized carbapenemase diagnostic assays.

Original language	English
Article number	114926
Journal	Diagnostic Microbiology and Infectious Disease
Volume	96
Issue number	4
DOIs	https://doi.org/10.1016/j.diagmicrobio.2019.114926
Publication status	Published - Apr 2020

Bibliographical note

Funding Information:
This study was funded by the National Institute for Health Research (NIHR) Invention for Innovation programme (Project reference, II-0511-21002; Rapid detection of infectious agents at point of triage (PoT). The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care.

Funding Information:
This study was funded by the National Institute for Health Research (NIHR) Invention for Innovation programme (Project reference, II-0511-21002; Rapid detection of infectious agents at point of triage (PoT). The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care. The funding source had no involvement in the design of the study, in the collection, analysis and interpretation of data, in the writing of the report or in the decision to submit the article for publication. All decisions related to submission of the manuscript were made by the author(s). We thank the Public Health England, Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit for supplying panels of carbapenemase-producing isolates for evaluation.

Publisher Copyright:
© 2019

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10.1016/j.diagmicrobio.2019.114926

Cite this

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title = "Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin",

abstract = "Routine identification of carbapenemase-producing bacterial isolates is a lengthy process often taking up to 72 h to generate results with standard culture-based tests. Here we describe a rapid test based on the hydrolysis of nitrocefin to identify isolates producing β-lactamase enzymes. A cocktail of inhibitors has been optimized in the reaction mix to provide specificity for carbapenemase enzymes. The developed assay has also been translated to a microfluidic platform with an optical readout (optofluidic chip). The chip has a long absorbance path (25 mm) to provide high sensitivity. A sample-to-answer has been achieved in under 30 min on these chips using colonies from culture plates. The test on this platform has the potential to provide a rapid indicative (presumptive positive) test for carbapenemase producers direct from bacteria isolated from patient samples, to rapidly trigger infection control measures and identify samples that should be prioritized for more specialized carbapenemase diagnostic assays.",

author = "Collette Allen and Carrie Turner and Sumit Kalsi and David Jamieson and Yuetao Li and Hywel Morgan and John Sutton",

note = "Funding Information: This study was funded by the National Institute for Health Research (NIHR) Invention for Innovation programme (Project reference, II-0511-21002; Rapid detection of infectious agents at point of triage (PoT). The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care. Funding Information: This study was funded by the National Institute for Health Research (NIHR) Invention for Innovation programme (Project reference, II-0511-21002; Rapid detection of infectious agents at point of triage (PoT). The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care. The funding source had no involvement in the design of the study, in the collection, analysis and interpretation of data, in the writing of the report or in the decision to submit the article for publication. All decisions related to submission of the manuscript were made by the author(s). We thank the Public Health England, Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit for supplying panels of carbapenemase-producing isolates for evaluation. Publisher Copyright: {\textcopyright} 2019",

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AU - Morgan, Hywel

AU - Sutton, John

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Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin

Abstract

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