Single Electrode Genosensor for Simultaneous Determination of Sequences Encoding Hemagglutinin and Neuraminidase of Avian Influenza Virus Type H5N1
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- 作者：Iwona Grabowska ; Kamila Malecka ; Anna Stachyra ; Anna G贸ra-Sochacka ; Agnieszka Sirko ; W艂odzimierz Zag贸rski-Ostoja ; Hanna Radecka ; Jerzy Radecki
- 刊名：Analytical Chemistry
- 出版年：2013
- 出版时间：November 5, 2013
- 年：2013
- 卷：85
- 期：21
- 页码：10167-10173
- 全文大小：317K
- 年卷期：v.85,no.21(November 5, 2013)
- ISSN：1520-6882
文摘
The duo-genosensor consisting of two different oligonucleotide probes immobilized covalently on the surface of one gold electrode via Au鈥揝 bond formation was used for simultaneous determination of two different oligonucleotide targets. One of the probes, decorated on its 5鈥?end with ferrocene (SH-ssDNA-Fc), is complementary to the cDNA representing a sequence encoding part of H5 hemagglutinin from H5N1 virus. The second probe, decorated on its 5鈥?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 the hybridization process were used as analytical signal. The peak at +400 卤 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 鈥渟ignal on鈥?mode of analytical signal generation. The peak at 鈭?50 卤 4 mV, characteristic for SH-ssDNA-MB probe, was decreasing with the increase of the concentration of the complementary ssDNA target starting from 8 to 100 nM. This indicates the generation of electrochemical signal according to the 鈥渟ignal off鈥?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.