Sequences of Pandemic-Causing Viruses Isolated and Detected by Paramagnetic Particles Coupled with Microfluidic System and Electrochemical Detector

摘要

Detection of the specific sequences of pandemic-causing viral nucleic acids is area attracting numerousscientists worldwide. In this study, we focused on the isolation and detection of oligonucleotidespecific sequences of life-threatening viruses (HIV (GAGCAGTGGGAATA), Influenza(TAATAACCATTGGA) and Ebola (ACCTCACTAGAAAA)) by flow injection analysis withelectrochemical detection coupled with paramagnetic particles based isolation. All of used viralgenomic sequences contained A6 adenine repetition on the 5end. This modification enables us to bindthem on paramagnetic particle anchoring T25 (MPs-(dT)25). The isolated sequences were subsequentlyhydrolysed by optimized method using microwaves to obtain single adenines. The obtained sampleswere detected by microfluidic tool based on stopped flow injection analysis with glassy carbonelectrode in the presence of Cu(II)/or CuS nanoparticles to enhance sensitivity and to lower detectionlimit for adenine. Basic detection parameters as accumulation of CuS nanoparticles, flow influence andaccumulation of the adenine deposition influencing the analytical response were also tested. We foundthat bases gave well developed signals with detection limit down to 30 ng per ml (50 pg perelectrochemical cell) under the optimized conditions.