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Programmable digital microfluidic assay for simultaneous detection of multiple anti-microbial resistance genes

Programmable digital microfluidic assay for simultaneous detection of multiple anti-microbial resistance genes
Programmable digital microfluidic assay for simultaneous detection of multiple anti-microbial resistance genes
The rapid emergence of antimicrobial resistant bacteria requires the development of new diagnostic tests. Nucleic acid based assays determine antimicrobial susceptibility by detecting genes that encode for the resistance. In this study, we demonstrate rapid and simultaneous detection of three genes that confer resistance in bacteria to extended spectrum β-lactam and carbapenem antibiotics; CTX-M-15, KPC and NDM-1. The assay uses isothermal DNA amplification (Recombinase Polymerase Amplification, RPA) implemented on a programmable digital microfluidics (DMF) platform. Automated dispensing protocols are used to simultaneously manipulate 45 droplets of nL volume containing sample DNA, reagents and controls. The droplets are processed and mixed under electronic control on the DMF devices with positive amplification measured by fluorescence. The assay on these devices is significantly improved with a Time to Positivity (TTP) twice that of the benchtop assay.
Kalsi, Sumit
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Sellars, Samuel, Lee
3a7253a9-c854-4b18-ad56-9d2e95e3467c
Turner, Carrie
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Sutton, Mark
d533d794-3525-4433-a973-af499e0cb93a
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Kalsi, Sumit
0ec1f5c2-a6ec-4c07-92fb-15708c28742e
Sellars, Samuel, Lee
3a7253a9-c854-4b18-ad56-9d2e95e3467c
Turner, Carrie
8ff2a882-4649-4031-a660-7b5889169557
Sutton, Mark
d533d794-3525-4433-a973-af499e0cb93a
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Kalsi, Sumit, Sellars, Samuel, Lee, Turner, Carrie, Sutton, Mark and Morgan, Hywel (2017) Programmable digital microfluidic assay for simultaneous detection of multiple anti-microbial resistance genes. Micromachines, 8 (4). (doi:10.3390/mi8040111).

Record type: Article

Abstract

The rapid emergence of antimicrobial resistant bacteria requires the development of new diagnostic tests. Nucleic acid based assays determine antimicrobial susceptibility by detecting genes that encode for the resistance. In this study, we demonstrate rapid and simultaneous detection of three genes that confer resistance in bacteria to extended spectrum β-lactam and carbapenem antibiotics; CTX-M-15, KPC and NDM-1. The assay uses isothermal DNA amplification (Recombinase Polymerase Amplification, RPA) implemented on a programmable digital microfluidics (DMF) platform. Automated dispensing protocols are used to simultaneously manipulate 45 droplets of nL volume containing sample DNA, reagents and controls. The droplets are processed and mixed under electronic control on the DMF devices with positive amplification measured by fluorescence. The assay on these devices is significantly improved with a Time to Positivity (TTP) twice that of the benchtop assay.

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More information

Accepted/In Press date: 7 March 2017
e-pub ahead of print date: 1 April 2017
Published date: April 2017

Identifiers

Local EPrints ID: 412507
URI: https://eprints.soton.ac.uk/id/eprint/412507
PURE UUID: b5136b6a-f1e7-4edb-8873-0f832efb806b
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 17 Jul 2017 14:01
Last modified: 24 Sep 2019 05:05

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