title: New redox-active layer create via epoxy-amine reaction - The base of genosensor for the detection of specific DNA and RNA sequences of avian influenza virus H5N1.
abstract: This paper concerns the development of a redox-active monolayer and its application for the construction of an electrochemical genosensor designed for the detection of specific DNA and RNA oligonucleotide sequences related to the avian influenza virus (AIV) type H5N1. This new redox layer was created on a gold electrode surface step by step. Cyclic Voltammetry, Osteryoung Square-Wave Voltammetry and Differential Pulse Voltammetry were used for its characterization. This new redox-active layer was applied for the construction of the DNA biosensor. The NH2-NC3 probe (20-mer) was covalently attached to the gold electrode surface via a "click" reaction between the amine and an epoxide group. The hybridization process was monitored using the Osteryoung Square-Wave Voltammetry. The 20-mer DNA and ca. 280-mer RNA oligonucleotides were used as the targets. The constructed genosensor was capable to determine complementary oligonucleotide sequences with a detection limit in the pM range. It is able to distinguish the different position of the part RNA complementary to the DNA probe. The genosensor was very selective. The 20-mer DNA as well as the 280-mer RNA oligonucleotides without a complementary sequence generated a weak signal.
authors.affiliation: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Malecka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Stachyra
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Góra-Sochacka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Sirko
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland.
authors.forename: Włodzimierz
authors.initials: W
authors.lastname: Zagórski-Ostoja
authors.affiliation: University of Leuven, Department of Chemistry, Celestijnenlaan 200F, B-3001 Leuven, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Dehaen
authors.affiliation: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
authors.forename: Hanna
authors.initials: H
authors.lastname: Radecka
authors.affiliation: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland. Electronic address: j.radecki@pan.olsztyn.pl.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Radecki
mesh_terms: D000586:Amination; D000818:Animals; D015374:Biosensing Techniques; D001717:Birds / Q000821:virology*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D004852:Epoxy Compounds / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D009693:Nucleic Acid Hybridization / Q000379:methods; D010084:Oxidation-Reduction; D010618:Phenanthrolines / Q000737:chemistry; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics
chemical_list: D015342:DNA Probes; D004852:Epoxy Compounds; D010618:Phenanthrolines; D012367:RNA, Viral; D006046:Gold
keywords: Avian influenza virus; Electrochemical genosensor; Epoxide–amine “click”; Phenanthroline-Epoxy–Fe(III)–(Phenanthroline Epoxy–NH(2)-ssDNA)(2) redox active layer; RNA transcripts