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Ultrasensitive electrochemical genosensor for direct detection of specific RNA sequences derived from avian influenza viruses present in biological samples

Malecka, Kamila and Świętoń, Edyta and Verwilst, Peter and Stachyra, Anna and Sirko, Agnieszka and Dehaen, Wim and Radecki, Jerzy and Radecka, Hanna (2019) Ultrasensitive electrochemical genosensor for direct detection of specific RNA sequences derived from avian influenza viruses present in biological samples. Acta Biochimica Polonica, 66 (3). pp. 299-304. ISSN 1734-154X

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Official URL: http://www.actabp.pl/

Abstract

An electrochemical genosensor based on an epoxyphenanthroline–Fe(III)–NH2-ssDNA layer for the detection of RNA derived from Avian Influenza is presented. The biosensor preparation consists of: (I) modification of gold electrodes with aminoethanethiol, (II) modification of the self-assembled monolayer of aminoethanethiol with 5,6-epoxy-5,6-dihydro-[1,10]-phenanthroline using “click” chemistry, (III) a first step of complexation of Fe(III) by 5,6-epoxy-5,6-dihydro-[1,10]-phenanthroline, (IV) a second step of complexation of Fe(III) by 5,6-epoxy-5,6-dihydro-[1,10]-phenanthroline, (V) immobilization of the single stranded amino-DNA probe via “click” chemistry between epoxy and amino groups. The interactions between the ssDNA probe and RNA targets were explored with Osteryoung Square Wave Voltammetry. The genosensor showed a remarkable detection limit of 3 copies/μL (5 aM) for RNA extracted from A/swan/Poland/305/06 (H5N1) containing a fully complementary sequence. A linear dynamic range for this sequence was observed from 3.0×103 to 3.0×105 [copies/μl]. RNA extracted from A/mallard/Poland/446/09 (H7N7), containing a non-complementary sequence, generated a much weaker response. Moreover, the developed genosensor allows to distinguish RNA present in biological samples having 2, 3 and 4 mismatches. This biosensing approach can become a potential alternative tool for detecting RNA samples in biomedical research and early clinical diagnosis of avian influenza viruses.

Item Type:Article
Subjects:Q Science > QD Chemistry
Q Science > QR Microbiology > QR355 Virology
Divisions:Department of Plant Biochemistry
Department of Protein Biosynthesis
ID Code:1754
Deposited By: Agnieszka Sirko
Deposited On:04 Oct 2019 12:39
Last Modified:04 Oct 2019 12:39

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