Simultaneous single-channel multiplexing and quantification of carbapenem-resistant genes using multidimensional standard curves

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Title: Simultaneous single-channel multiplexing and quantification of carbapenem-resistant genes using multidimensional standard curves
Authors: Rodriguez-Manzano, J
Moniri, A
Malpartida-Cardenas, K
Dronavalli, J
Davies, F
Holmes, A
Georgiou, P
Item Type: Journal Article
Abstract: Multiplexing and quantification of nucleic acids, both have, in their own right, significant and extensive use in biomedical related fields. Currently, the ability to detect several nucleic acid targets in a single-reaction scales linearly with the number of targets; an expensive and time-consuming feat. Here, we propose a new methodology based on multidimensional standard curves that extends the use of real-time PCR data obtained by common qPCR instruments. By applying this novel method-ology, we achieve simultaneous single-channel multiplexing and enhanced quantification of multiple targets using only real-time amplification data. This is obtained without the need of fluorescent probes, agarose gels, melting curves or sequencing analysis. Given the importance and demand for tackling challenges in antimicrobial resistance, the proposed method is ap-plied to four of the most prominent carbapenem-resistant genes: blaOXA-48, blaNDM, blaVIM and blaKPC, which account for 97% of the UK's reported carbapenemase-producing Enterobacteriaceae.
Issue Date: 5-Feb-2019
Date of Acceptance: 9-Jan-2019
URI: http://hdl.handle.net/10044/1/65997
DOI: https://d.xdoi.org/10.1021/acs.analchem.8b04412
ISSN: 0003-2700
Publisher: American Chemical Society
Start Page: 2013
End Page: 2020
Journal / Book Title: Analytical Chemistry
Volume: 91
Issue: 3
Copyright Statement: © 2019 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, after peer review and technical editing by the publisher. To access the final edited and published work see https://dx.doi.org/10.1021/acs.analchem.8b04412
Sponsor/Funder: National Institute for Health Research
National Institute for Health Research
Engineering & Physical Science Research Council (EPSRC)
Funder's Grant Number: HPRU-2012-10047
HPRU-2012-10047
EP/R512655/1
Keywords: 0301 Analytical Chemistry
0904 Chemical Engineering
0399 Other Chemical Sciences
Analytical Chemistry
Publication Status: Published
Conference Place: United States
Embargo Date: 2020-01-09
Online Publication Date: 2019-01-09
Appears in Collections:Faculty of Engineering
Electrical and Electronic Engineering
Department of Medicine
Faculty of Medicine



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