tetano
Editor, Senior Moderator
Anal Chem. 2013 Sep 25. [Epub ahead of print]
Single Electrode Genosensor for Simultaneous Determination of Sequences Encoding Hemagglutinin and Neuraminidase of Avian Influenza Virus (AIV) type H5N1.
Grabowska I, Malecka K, Stachyra A, G?ra-Sochacka A, Sirko A, Zag?rski-Ostoja W, Radecka H, Radecki J.
Abstract
The duo-genosensor consisting of two different oligonucleotide probes immobilized covalently on the surface of one gold electrode via Au-S bond formation was used for simultaneous determination of two different oligonucleotide targets. One of the probes, decorated on its 5'-end with ferrocene (SH-ssDNA-Fc), is complementary to the cDNA representing sequence encoding part of H5 hemagglutinin from H5N1 virus. The second probe, decorated on its 5'-end with methylene blue (SH-ssDNA-MB), is complementary to cDNA representing the fragment of N1 neuraminidase from the same virus. The presence of both probes on the surface of gold electrodes was confirmed with Osteryoung square-wave voltammetry (OSWV). The changes in redox activity of both redox active complexes before and after hybridization process were used as analytical signal. The peak at +400 2 mV was observed in the presence of 40 nM ssDNA used as a target for SH-ssDNA-Fc probe. This peak increased with the increase of concentration of target ssDNA. It indicates the "signal on" mode of analytical signal generation. The peak at -250 4 mV, characteristic for SH-ssDNA-MB probe was decreasing with the increase of the concentration of the complementary ssDNA target starting from 8 nM to 100 nM. This indicates the generation of electrochemical signal according to the "signal off" mode. The proposed duo-genosensor is capable of simultaneous, specific and good sensitivity probing for the sequences derived from genes encoding two main markers of the influenza virus, hemagglutinin and neuraminidase.
PMID:
24063372
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24063372
Single Electrode Genosensor for Simultaneous Determination of Sequences Encoding Hemagglutinin and Neuraminidase of Avian Influenza Virus (AIV) type H5N1.
Grabowska I, Malecka K, Stachyra A, G?ra-Sochacka A, Sirko A, Zag?rski-Ostoja W, Radecka H, Radecki J.
Abstract
The duo-genosensor consisting of two different oligonucleotide probes immobilized covalently on the surface of one gold electrode via Au-S bond formation was used for simultaneous determination of two different oligonucleotide targets. One of the probes, decorated on its 5'-end with ferrocene (SH-ssDNA-Fc), is complementary to the cDNA representing sequence encoding part of H5 hemagglutinin from H5N1 virus. The second probe, decorated on its 5'-end with methylene blue (SH-ssDNA-MB), is complementary to cDNA representing the fragment of N1 neuraminidase from the same virus. The presence of both probes on the surface of gold electrodes was confirmed with Osteryoung square-wave voltammetry (OSWV). The changes in redox activity of both redox active complexes before and after hybridization process were used as analytical signal. The peak at +400 2 mV was observed in the presence of 40 nM ssDNA used as a target for SH-ssDNA-Fc probe. This peak increased with the increase of concentration of target ssDNA. It indicates the "signal on" mode of analytical signal generation. The peak at -250 4 mV, characteristic for SH-ssDNA-MB probe was decreasing with the increase of the concentration of the complementary ssDNA target starting from 8 nM to 100 nM. This indicates the generation of electrochemical signal according to the "signal off" mode. The proposed duo-genosensor is capable of simultaneous, specific and good sensitivity probing for the sequences derived from genes encoding two main markers of the influenza virus, hemagglutinin and neuraminidase.
PMID:
24063372
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24063372