The optimization of an electrochemical aptasensor to detect RBD protein S SARS-CoV-2 as a biomarker of COVID-19 using screen-printed carbon electrode/AuNP

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus identified as the cause of the Coronavirus Disease 2019 (COVID-19) outbreak. The gold standard for detecting this virus is polymerase chain reaction (PCR). The electrochemical biosensor method can be an alternative method for detecting several biomolecules, such as viruses, because it is proven to have several advantages, including portability, good sensitivity, high specificity, fast response, and ease of use. This study aims to optimize an electro­chemical aptasensor using an AuNP-modified screen-printed carbon electrode (SPCE) with an aptamer to detect RBD protein S SARS-CoV-2. Aptasensors with the streptavidin-biotin system were immobilized on the SPCE/AuNP surface via covalent bonding with linkers to 3-mercaptopropionic acid (MPA) and electrochemically characterized by the [Fe(CN)6]3-/4- redox system using differential pulse voltammetry. The results showed that the immobi­lized aptamer on the SPCE/AuNP electrode surface experienced a decrease in current from 11.388 to 4.623 µA. The experimental conditions were optimized using the Box-Behnken experimental design for the three factors that affect the current response. The results of the optimization of the three parameters are the concentration of aptamer, incubation time of aptamer, and incubation time of RBD protein S SARS-CoV-2, each of which is 0.5 µg/mL, 40 minutes, and 60 minutes, respectively. The RBD protein S SARS-CoV-2 with various concentrations was tested on an electrochemical aptasensor to determine the de­tection limit and quantification limit, and the respective results were 2.63 and 7.97 ng/mL. The electrochemical aptasensor that has been developed in this study can be applied to detect RBD protein S SARS-CoV-2 as a COVID-19 biomarker in a simple, practical, and sensitive way.