title: DNA probe modified with 3-iron bis(dicarbollide) for electrochemical determination of DNA sequence of Avian Influenza Virus H5N1.
abstract: In this work, we report on oligonucleotide probes bearing metallacarborane [3-iron bis(dicarbollide)] redox label, deposited on gold electrode for electrochemical determination of DNA sequence derived from Avian Influenza Virus (AIV), type H5N1. The oligonucleotide probes containing 5'-terminal NH2 group were covalently attached to the electrode, via NHS/EDC coupling to 3-mercaptopropionic acid SAM, previously deposited on the surface of gold. The changes in redox activity of Fe(III) centre of the metallacarborane complex before and after hybridization process was used as analytical signal. The signals generated upon hybridization with targets such as complementary or non-complementary 20-mer ssDNA or various PCR products consisting of 180-190 bp (dsDNA) were recorded by Osteryoung square-wave voltammetry (OSWV). The developed system was very sensitive towards targets containing sequence complementary to the probe with the detection limit estimated as 0.03 fM (S/N=3.0) and 0.08 fM (S/N=3.0) for 20-mer ssDNA and for dsDNA (PCR product), respectively. The non-complementary targets generated very weak responses. Furthermore, the proposed genosensor was suitable for discrimination of PCR products with different location of the complementarity region.
authors: Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
authors: Iwona
authors: I
authors: Grabowska
authors: Anna
authors: A
authors: Stachyra
authors: Anna
authors: A
authors: Góra-Sochacka
authors: Agnieszka
authors: A
authors: Sirko
authors: Agnieszka B
authors: AB
authors: Olejniczak
authors: Zbigniew J
authors: ZJ
authors: Leśnikowski
authors: Jerzy
authors: J
authors: Radecki
authors: Hanna
authors: H
authors: Radecka
mesh_terms: D000818:Animals; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods; D001717:Birds / Q000821:virology; D015342:DNA Probes / Q000737:chemistry*; D004279:DNA, Viral / Q000032:analysis*; D055664:Electrochemical Techniques / Q000379:methods; D005290:Ferric Compounds / Q000737:chemistry*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology*; D057230:Limit of Detection; D008958:Models, Molecular; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010084:Oxidation-Reduction
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D005290:Ferric Compounds
keywords: 3-Iron bis(dicarbollide)-oligonucleotide; Avian Influenza Virus (AIV); Electrochemical ssDNA detection; Osteryoung square-wave voltammetry