Electrochemical detection of oxacillin resistance using direct-labelling solid-phase isothermal amplification
Butterworth, Adrian and Pratibha, Pratibha and Marx, Andreas and Corrigan, Damion K. (2021) Electrochemical detection of oxacillin resistance using direct-labelling solid-phase isothermal amplification. ACS Sensors, 6 (10). pp. 3773-3780. ISSN 2379-3694 (https://doi.org/10.1021/acssensors.1c01688)
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Abstract
Isothermal amplification reactions represent an important and exciting approach to achieve widespread, low cost, and easily implemented molecular diagnostics. This work presents a modified recombinase polymerase amplification (RPA) reaction, which can be directly coupled to a simple electrochemical measurement to ultimately allow development of a nucleic acid-based assay for antibiotic resistance genes. It is shown that use of reagents from a standard RPA reaction kit allows incorporation of horse radish peroxidase-labeled thymine nucleotides into amplified DNA strands, which can be detected via an amperometric signal readout for detection of important gene sequences. The assay is exemplified through detection of fragments of the oxacillin resistance gene in Escherichia coli cells bearing a drug resistance plasmid, achieving a potential limit of detection of 319 cfus/mL and an unoptimized time to result of 60 min. This work serves as a suitable demonstration of the potential for a system to deliver detection of key drug resistance genes at clinically relevant levels.
ORCID iDs
Butterworth, Adrian ORCID: https://orcid.org/0000-0002-0463-927X, Pratibha, Pratibha, Marx, Andreas and Corrigan, Damion K. ORCID: https://orcid.org/0000-0002-4647-7483;-
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Item type: Article ID code: 77985 Dates: DateEvent22 October 2021Published1 October 2021Published Online24 September 2021AcceptedSubjects: Technology > Chemical technology Department: Faculty of Engineering > Biomedical Engineering Depositing user: Pure Administrator Date deposited: 01 Oct 2021 15:27 Last modified: 11 Nov 2024 13:14 URI: https://strathprints.strath.ac.uk/id/eprint/77985