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Sample pre-concentration on a digital microfluidic platform for rapid AMR detection in urine

Sample pre-concentration on a digital microfluidic platform for rapid AMR detection in urine
Sample pre-concentration on a digital microfluidic platform for rapid AMR detection in urine
There is a growing need for rapid diagnostic methods to support stewardship of antibiotics. We describe an analytical platform for sample concentration to detect antimicrobial resistance (AMR) genes directly from human urine for the diagnosis of Urinary Tract Infections (UTIs) that are resistant to antibiotics. A sample-processing unit concentrates plasmid DNA directly from urine using magnetic beads, followed by isothermal amplification of target genes. The sample pre-concentration unit interfaces with a digital microfluidic platform (DMF) and scales the sample volume by 500-fold, pre-concentrating DNA from 1mL into a 2L droplet for downstream processing. Tests with a clinical strain of Klebsiella pneumoniae (NCTC 13443), spiked into human urine demonstrated a limit of detection of 104 cfu/mL and a “sample to answer” in approximately 30 minutes.
1473-0197
168-177
Kalsi, Sumit
0ec1f5c2-a6ec-4c07-92fb-15708c28742e
Valiadi, Martha
e245e9c5-3e97-4896-b910-c4178c5fb3c9
Turner, Carrie
8ff2a882-4649-4031-a660-7b5889169557
Sutton, J. Mark
d533d794-3525-4433-a973-af499e0cb93a
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Kalsi, Sumit
0ec1f5c2-a6ec-4c07-92fb-15708c28742e
Valiadi, Martha
e245e9c5-3e97-4896-b910-c4178c5fb3c9
Turner, Carrie
8ff2a882-4649-4031-a660-7b5889169557
Sutton, J. Mark
d533d794-3525-4433-a973-af499e0cb93a
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Kalsi, Sumit, Valiadi, Martha, Turner, Carrie, Sutton, J. Mark and Morgan, Hywel (2019) Sample pre-concentration on a digital microfluidic platform for rapid AMR detection in urine. Lab on a Chip, 19, 168-177. (doi:10.1039/C8LC01249K).

Record type: Article

Abstract

There is a growing need for rapid diagnostic methods to support stewardship of antibiotics. We describe an analytical platform for sample concentration to detect antimicrobial resistance (AMR) genes directly from human urine for the diagnosis of Urinary Tract Infections (UTIs) that are resistant to antibiotics. A sample-processing unit concentrates plasmid DNA directly from urine using magnetic beads, followed by isothermal amplification of target genes. The sample pre-concentration unit interfaces with a digital microfluidic platform (DMF) and scales the sample volume by 500-fold, pre-concentrating DNA from 1mL into a 2L droplet for downstream processing. Tests with a clinical strain of Klebsiella pneumoniae (NCTC 13443), spiked into human urine demonstrated a limit of detection of 104 cfu/mL and a “sample to answer” in approximately 30 minutes.

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c8lc01249k-1 - Accepted Manuscript
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More information

Accepted/In Press date: 22 November 2018
e-pub ahead of print date: 23 November 2018
Published date: 7 January 2019

Identifiers

Local EPrints ID: 426415
URI: http://eprints.soton.ac.uk/id/eprint/426415
ISSN: 1473-0197
PURE UUID: 8160a0d5-485f-4a07-ad9a-9d6c61e4131b
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

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Date deposited: 27 Nov 2018 17:30
Last modified: 16 Mar 2024 07:20

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Contributors

Author: Sumit Kalsi
Author: Martha Valiadi
Author: Carrie Turner
Author: J. Mark Sutton
Author: Hywel Morgan ORCID iD

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