Publication details
ISOLATION AND VOLTAMMETRY DETECTION OF GENOME SEQUENCE EQUINE INFLUENZA VIRUS SUBTYPE 2 (H3N8) USING MAGNETIC PARTICLES
| Authors | |
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| Year of publication | 2011 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Equine influenza virus subtype 2 (H3N8) is a causal agent of respiratory disease of equids. The aim of this study was to design detection system that enables reliable isolation of this pathogen at low concentration levels. To isolate specific viral nucleic acid streptavidin-coupled paramagnetic nano- and microparticles were employed and optimized. Biotin-labelled DNA probes were targeted to four genes encoding neuraminidase, nucleoprotein, hemaglutinin and matrix proteins of H3N8 virus. The concentration of extracted viral oligonucleotide sequence was measured by using square wave voltammetry at hanging mercury drop electrode. Thus, the experimental conditions were optimized. The optimized parameters were as follows – supporting electrolyte: acetate buffer pH 5, frequency: 280 Hz, step potential: 5 mV and amplitude 25 mV. Under these conditions, the concentration range for all tested viral oligonucleotide genes was from 0.1 to 3 ng/microl. We determined four linear calibration curves with R higher than 0.99. The detection limits for all four genes were estimated down to tens of pg per microl. It clearly follows from the results obtained that coupling of paramagnetic particles and electrochemical detection is very promising in DNA analysis with respect to great potential in miniaturization. |