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Detection of PCR-Amplified Tuberculosis DNA Fragments with Polyelectrolyte-Modified Field-Effect Sensors

  • Field-effect-based electrolyte-insulator-semiconductor (EIS) sensors were modified with a bilayer of positively charged weak polyelectrolyte (poly(allylamine hydrochloride) (PAH)) and probe single-stranded DNA (ssDNA) and are used for the detection of complementary single-stranded target DNA (cDNA) in different test solutions. The sensing mechanism is based on the detection of the intrinsic molecular charge of target cDNA molecules after the hybridization event between cDNA and immobilized probe ssDNA. The test solutions contain synthetic cDNA oligonucleotides (with a sequence of tuberculosis mycobacteria genome) or PCR-amplified DNA (which origins from a template DNA strand that has been extracted from Mycobacterium avium paratuberculosis-spiked human sputum samples), respectively. Sensor responses up to 41 mV have been measured for the test solutions with DNA, while only small signals of ∼5 mV were detected for solutions without DNA. The lower detection limit of the EIS sensors was ∼0.3 nM, and the sensitivity was ∼7.2 mV/decade. Fluorescence experiments using SybrGreen I fluorescence dye support the electrochemical results.

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Metadaten
Author:Thomas BronderORCiD, Max P. Jessing, Arshak PoghossianORCiD, Michael Keusgen, Michael Josef SchöningORCiD
DOI:https://doi.org/10.1021/acs.analchem.8b01807
ISSN:0003-2700
Parent Title (English):Analytical Chemistry
Publisher:ACS Publications
Place of publication:Washington, DC
Document Type:Article
Language:English
Year of Completion:2018
Date of the Publication (Server):2018/07/04
Volume:90
Issue:12
First Page:7747
Last Page:7753
Link:https://doi.org/10.1021/acs.analchem.8b01807
Zugriffsart:campus
Institutes:FH Aachen / Fachbereich Medizintechnik und Technomathematik
FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / ACS Publications