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Cystic fibrosis: a label-free detection approach based on thermally modulated electrochemical impedance spectroscopy
Universitat Rovira i Virgili, Tarragona, Spain.
Universitat Rovira i Virgili, Tarragona, Spain.ORCID iD: 0000-0001-6889-0351
Universitat Rovira i Virgili, Tarragona, Spain.
Universitat Rovira i Virgili, Tarragona, Spain.
2010 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 396, no 7, 2565-2574 p.Article in journal (Refereed) Published
Abstract [en]

Cystic fibrosis is one of the most commongenetically inherited diseases in northern Europe, with theDF508 mutation being the most common, and among theCaucasian population being responsible for almost 70% ofcases. In this work, we report on the use of thermallymodulated electrochemical impedance spectroscopy for thediscrimination of the DF508 mutation from the wild-typesequence. DNA probes (15 and 21 bases long) wereimmobilised on the surface of gold electrodes and thevariation of the charge-transfer resistance was monitored asa function of hybridisation. Two sets of targets were used inthis work: synthetic 15-mer sequences and two singlestrandedsynthetic analogues of PCR products 82 (mutant)and 85 (wild type) bases long. Hybridisation with shorttargets resulted in very sequence specific charge-transferresistancevariation with a discrimination factor at roomtemperature between fully complementary and mismatchedsequences of approximately fivefold. However, in the caseof the single-stranded synthetic PCR product analogues, alower discrimination factor was recorded (1.5-fold). Theeffect of temperature was investigated to improve discriminationand the use of a posthybridisation wash at elevatedtemperature resulted in a fivefold improvement in thediscrimination factor. Using an electrode array with probesimmobilised against each of the mutant and wild-typesequences, we achieved an unequivocal detection of theDF508 mutation.

Place, publisher, year, edition, pages
Springer Publishing Company, 2010. Vol. 396, no 7, 2565-2574 p.
Keyword [en]
Electrochemical impedance spectroscopy . Cystic fibrosis . Label-less detection .DNA sensor . DF508
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:liu:diva-92112DOI: 10.1007/s00216-010-3489-yPubMedID: 20155413OAI: oai:DiVA.org:liu-92112DiVA: diva2:620161
Available from: 2013-05-07 Created: 2013-05-07 Last updated: 2017-12-06Bibliographically approved

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Beni, Valerio

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