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Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

It is well-known that two major issues, preventing improved outcomes from cancer are late diagnosis and the evolution of drug resistance during chemotherapy, therefore technologies that address these issues can have a transformative effect on healthcare workflows. In this work we present a simple, l...

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Autores principales: Attoye, Bukola, Pou, Chantevy, Blair, Ewen, Rinaldi, Christopher, Thomson, Fiona, Baker, Matthew J., Corrigan, Damion K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7692145/
https://www.ncbi.nlm.nih.gov/pubmed/33126531
http://dx.doi.org/10.3390/bios10110156
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author Attoye, Bukola
Pou, Chantevy
Blair, Ewen
Rinaldi, Christopher
Thomson, Fiona
Baker, Matthew J.
Corrigan, Damion K.
author_facet Attoye, Bukola
Pou, Chantevy
Blair, Ewen
Rinaldi, Christopher
Thomson, Fiona
Baker, Matthew J.
Corrigan, Damion K.
author_sort Attoye, Bukola
collection PubMed
description It is well-known that two major issues, preventing improved outcomes from cancer are late diagnosis and the evolution of drug resistance during chemotherapy, therefore technologies that address these issues can have a transformative effect on healthcare workflows. In this work we present a simple, low-cost DNA biosensor that was developed specifically to detect mutations in a key oncogene (KRAS). The sensor employed was a screen-printed array of carbon electrodes, used to perform parallel measurements of DNA hybridisation. A DNA amplification reaction was developed with primers for mutant and wild type KRAS sequences which amplified target sequences from representative clinical samples to detectable levels in as few as twenty cycles. High levels of sensitivity were demonstrated alongside a clear exemplar of assay specificity by showing the mutant KRAS sequence was detectable against a significant background of wild type DNA following amplification and hybridisation on the sensor surface. The time to result was found to be 3.5 h with considerable potential for optimisation through assay integration. This quick and versatile biosensor has the potential to be deployed in a low-cost, point-of-care test where patients can be screened either for early diagnosis purposes or monitoring of response to therapy.
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spelling pubmed-76921452020-11-28 Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids Attoye, Bukola Pou, Chantevy Blair, Ewen Rinaldi, Christopher Thomson, Fiona Baker, Matthew J. Corrigan, Damion K. Biosensors (Basel) Article It is well-known that two major issues, preventing improved outcomes from cancer are late diagnosis and the evolution of drug resistance during chemotherapy, therefore technologies that address these issues can have a transformative effect on healthcare workflows. In this work we present a simple, low-cost DNA biosensor that was developed specifically to detect mutations in a key oncogene (KRAS). The sensor employed was a screen-printed array of carbon electrodes, used to perform parallel measurements of DNA hybridisation. A DNA amplification reaction was developed with primers for mutant and wild type KRAS sequences which amplified target sequences from representative clinical samples to detectable levels in as few as twenty cycles. High levels of sensitivity were demonstrated alongside a clear exemplar of assay specificity by showing the mutant KRAS sequence was detectable against a significant background of wild type DNA following amplification and hybridisation on the sensor surface. The time to result was found to be 3.5 h with considerable potential for optimisation through assay integration. This quick and versatile biosensor has the potential to be deployed in a low-cost, point-of-care test where patients can be screened either for early diagnosis purposes or monitoring of response to therapy. MDPI 2020-10-28 /pmc/articles/PMC7692145/ /pubmed/33126531 http://dx.doi.org/10.3390/bios10110156 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Attoye, Bukola
Pou, Chantevy
Blair, Ewen
Rinaldi, Christopher
Thomson, Fiona
Baker, Matthew J.
Corrigan, Damion K.
Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title_full Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title_fullStr Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title_full_unstemmed Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title_short Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids
title_sort developing a low-cost, simple-to-use electrochemical sensor for the detection of circulating tumour dna in human fluids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7692145/
https://www.ncbi.nlm.nih.gov/pubmed/33126531
http://dx.doi.org/10.3390/bios10110156
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