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Simple and rapid sample preparation system for the molecular detection of antibiotic resistant bacteria in human urine

Simple and rapid sample preparation system for the molecular detection of antibiotic resistant bacteria in human urine
Simple and rapid sample preparation system for the molecular detection of antibiotic resistant bacteria in human urine
Antibiotic resistance in urinary tract infections (UTIs) can cause significant complications without quick detection and appropriate treatment. . We describe a new approach to capture, concentrate and prepare amplification-ready DNA from antibiotic resistant bacteria in human urine samples. Klebsiella pneumoniae NCTC13443 (blaCTX-M-15 positive) spiked into filtered human urine was used as a model system. Bacteria were captured using anion exchange diaethylaminoethyl (DEAE) magnetic microparticles and concentrated 200-fold within ~3.5 minutes using a custom, valve-less microfluidic chip. Eight samples were processed in parallel, and DNA, was released using heat lysis from an integrated resistive heater. The crude cell lysate was used for real time Recombinase Polymerase Amplification (RPA) of the blaCTX-M-15 gene. The end to end processing time was approximately 15 minutes with a limit of detection of 1000 bacteria in 1 mL urine
1387-2176
1-30
Valiadi, Martha
e245e9c5-3e97-4896-b910-c4178c5fb3c9
Kalsi, Sumit
65cb3d4c-c28c-490a-85a3-7a8fb96ecfee
Jones, Isaac
f550a9db-f6fd-43cb-825c-bbbaee87cf88
Turner, Carrie
92e6c17b-41e4-4867-9812-677e4cb88996
Sutton, Mark
3ffaeca5-caf0-4d77-a573-fa4763387149
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Valiadi, Martha
e245e9c5-3e97-4896-b910-c4178c5fb3c9
Kalsi, Sumit
65cb3d4c-c28c-490a-85a3-7a8fb96ecfee
Jones, Isaac
f550a9db-f6fd-43cb-825c-bbbaee87cf88
Turner, Carrie
92e6c17b-41e4-4867-9812-677e4cb88996
Sutton, Mark
3ffaeca5-caf0-4d77-a573-fa4763387149
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Valiadi, Martha, Kalsi, Sumit, Jones, Isaac, Turner, Carrie, Sutton, Mark and Morgan, Hywel (2016) Simple and rapid sample preparation system for the molecular detection of antibiotic resistant bacteria in human urine. Biomedical Microdevices, 1-30. (In Press)

Record type: Article

Abstract

Antibiotic resistance in urinary tract infections (UTIs) can cause significant complications without quick detection and appropriate treatment. . We describe a new approach to capture, concentrate and prepare amplification-ready DNA from antibiotic resistant bacteria in human urine samples. Klebsiella pneumoniae NCTC13443 (blaCTX-M-15 positive) spiked into filtered human urine was used as a model system. Bacteria were captured using anion exchange diaethylaminoethyl (DEAE) magnetic microparticles and concentrated 200-fold within ~3.5 minutes using a custom, valve-less microfluidic chip. Eight samples were processed in parallel, and DNA, was released using heat lysis from an integrated resistive heater. The crude cell lysate was used for real time Recombinase Polymerase Amplification (RPA) of the blaCTX-M-15 gene. The end to end processing time was approximately 15 minutes with a limit of detection of 1000 bacteria in 1 mL urine

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Valiadi et al submission Biomed Microdev.pdf - Accepted Manuscript
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More information

Accepted/In Press date: 5 January 2016
Organisations: Nanoelectronics and Nanotechnology

Identifiers

Local EPrints ID: 385553
URI: https://eprints.soton.ac.uk/id/eprint/385553
ISSN: 1387-2176
PURE UUID: 325aeea2-7701-4c3a-87ce-a93b9700850a
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 20 Jan 2016 10:27
Last modified: 06 Jun 2018 12:45

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