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Dataset for Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes

Dataset for Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes
Dataset for Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes
Dataset supports: Nakano, M. I., Kalsi, S., & Morgan, H. (2018). Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes. Biosensors & Bioelectronics.Antimicrobial resistant pathogens are a growing worldwide threat to human health. This study proposes a novel method for rapid and sensitive detection of antimicrobial resistance (AMR) genes, specifically blaCTX-M-15 which encodes for the enzyme that offers resistance to extended spectrum β-lactam antibiotics. The method combines isothermal DNA amplification by recombinase polymerase amplification (RPA), with microbead dielectrophoresis (DEP)-based DNA detection. The amplicon of RPA is attached to dielectric microbeads, and the amount of amplicon determined by dielectrophoretic impedance measurement (DEPIM) of the microbeads. Amplicon-labeled microbeads were prepared by two-step and one-step labeling. A purified recombinant plasmid containing blaCTX-M-15 and genomic DNA (with plasmid) extracted from AMR bacteria (Escherichia coli NCTC 13441) were used as target samples. A one-step method in which RPA and DNA immobilization on the microbeads are carried out simultaneously, has a detection limit of 2 copies/reaction for the pure plasmid and 50 copies/reaction for genomic DNA. The assays are quantitative with a dynamic range up to 105 copies/reaction, with a total detection time of 26 min. Both methods are easy, rapid, and unlike lateral flow detection are quantitative.
University of Southampton
Nakano, MIchihiko
fbbd1e17-7f3c-4d33-a716-18fddcdb7192
Kalsi, Sumit
0ec1f5c2-a6ec-4c07-92fb-15708c28742e
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Nakano, MIchihiko
fbbd1e17-7f3c-4d33-a716-18fddcdb7192
Kalsi, Sumit
0ec1f5c2-a6ec-4c07-92fb-15708c28742e
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Nakano, MIchihiko (2018) Dataset for Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes. University of Southampton doi:10.5258/SOTON/D0572 [Dataset]

Record type: Dataset

Abstract

Dataset supports: Nakano, M. I., Kalsi, S., & Morgan, H. (2018). Fast and sensitive isothermal DNA assay using micro-bead dielectrophoresis for detection of anti-microbial resistance genes. Biosensors & Bioelectronics.Antimicrobial resistant pathogens are a growing worldwide threat to human health. This study proposes a novel method for rapid and sensitive detection of antimicrobial resistance (AMR) genes, specifically blaCTX-M-15 which encodes for the enzyme that offers resistance to extended spectrum β-lactam antibiotics. The method combines isothermal DNA amplification by recombinase polymerase amplification (RPA), with microbead dielectrophoresis (DEP)-based DNA detection. The amplicon of RPA is attached to dielectric microbeads, and the amount of amplicon determined by dielectrophoretic impedance measurement (DEPIM) of the microbeads. Amplicon-labeled microbeads were prepared by two-step and one-step labeling. A purified recombinant plasmid containing blaCTX-M-15 and genomic DNA (with plasmid) extracted from AMR bacteria (Escherichia coli NCTC 13441) were used as target samples. A one-step method in which RPA and DNA immobilization on the microbeads are carried out simultaneously, has a detection limit of 2 copies/reaction for the pure plasmid and 50 copies/reaction for genomic DNA. The assays are quantitative with a dynamic range up to 105 copies/reaction, with a total detection time of 26 min. Both methods are easy, rapid, and unlike lateral flow detection are quantitative.

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Published date: 30 June 2018
Organisations: Biomedical Electronics, Institute for Life Sciences

Identifiers

Local EPrints ID: 421869
URI: https://eprints.soton.ac.uk/id/eprint/421869
PURE UUID: 5e75b95b-3d95-4c05-92c2-a80433719aaa
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 03 Jul 2018 16:31
Last modified: 05 Jul 2018 00:33

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Contributors

Creator: MIchihiko Nakano
Contributor: Sumit Kalsi
Contributor: Hywel Morgan ORCID iD

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