Abstract
We report an electrochemical biosensor combined with recombinase polymerase amplification (RPA) for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2. The electrochemical biosensor based on a multi-microelectrode array allows the detection of multiple target genes by differential pulse voltammetry. The RPA reaction involves hybridization of the RPA amplicon with thiol-modified primers immobilized on the working electrodes, which leads to a reduction of current density as amplicons accumulate. The assay results in shorter “sample-to-answer” times than conventional PCR without expensive thermo-cycling equipment. The limits of detection are about 0.972 fg/μL (RdRP gene) and 3.925 fg/μL (N gene), which are slightly lower than or comparable to that of RPA assay results obtained by gel electrophoresis without post-amplification purification. The combination of electrochemical biosensors and the RPA assay is a rapid, sensitive, and convenient platform that can be potentially used as a point-of-care test for the diagnosis of COVID-19.
Keywords: COVID-19; Coronavirus; Electrochemical detection method; Recombinase polymerase amplification; SARS-CoV-2.
【저자키워드】 COVID-19, SARS-CoV-2, coronavirus, recombinase polymerase amplification, Electrochemical detection method, 【초록키워드】 SARS-CoV-2, equipment, thiol, severe acute respiratory syndrome Coronavirus, point-of-care, amplification, PCR, RdRP, N gene, recombinase polymerase amplification, platform, RPA, Combination, RdRp gene, Amplicon, Limits of detection, target gene, reduction, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, limits, primer, pulse, hybridization, purification, polymerase, gel electrophoresis, amplicons, REcombinase, current density, comparable, accumulate, diagnosis of COVID-19, 【제목키워드】 point-of-care, isothermal amplification, sensitive, COVID-19 test,