tetano
Editor, Senior Moderator
Acta Biochim Pol. 2019 Aug 22. doi: 10.18388/abp.2019_2774. [Epub ahead of print]
[h=1]Ultrasensitive electrochemical genosensor for direct detection of specific RNA sequences derived from avian influenza viruses present in biological samples.[/h] Malecka K[SUP]1[/SUP], Świętoń E[SUP]2[/SUP], Verwilst P[SUP]3[/SUP], Stachyra A[SUP]4[/SUP], Sirko A[SUP]4[/SUP], Dehaen W[SUP]5[/SUP], Radecki J[SUP]1[/SUP], Radecka H[SUP]1[/SUP].
[h=3]Author information[/h] 1 Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland. 2 National Veterinary Research Institute, Puławy, Poland. 3 Korea University, Department of Chemistry, Seoul, South Korea. 4 Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland. 5 University of Leuven, Department of Chemistry, Leuven, Belgium.
[h=3]Abstract[/h] An electrochemical genosensor based on an epoxy-phenanthroline-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/?L (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/?l]. 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.
PMID: 31442009 DOI: 10.18388/abp.2019_2774
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[h=1]Ultrasensitive electrochemical genosensor for direct detection of specific RNA sequences derived from avian influenza viruses present in biological samples.[/h] Malecka K[SUP]1[/SUP], Świętoń E[SUP]2[/SUP], Verwilst P[SUP]3[/SUP], Stachyra A[SUP]4[/SUP], Sirko A[SUP]4[/SUP], Dehaen W[SUP]5[/SUP], Radecki J[SUP]1[/SUP], Radecka H[SUP]1[/SUP].
[h=3]Author information[/h] 1 Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland. 2 National Veterinary Research Institute, Puławy, Poland. 3 Korea University, Department of Chemistry, Seoul, South Korea. 4 Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland. 5 University of Leuven, Department of Chemistry, Leuven, Belgium.
[h=3]Abstract[/h] An electrochemical genosensor based on an epoxy-phenanthroline-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/?L (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/?l]. 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.
PMID: 31442009 DOI: 10.18388/abp.2019_2774
Free full text