Abstract
A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a cause of worldwide Coronavirus 2019 (COVID-19) disease pandemic. It is thus important to develop ultra-sensitive, rapid and easy-to-use methods for the identification of COVID-19 infected patients. Herein, an alternative electrochemical immunosensor based on poly(pyrrolepropionic acid) (pPPA) modified graphene screen-printed electrode (GSPE) was proposed for rapid COVID-19 detection. The method was based on a competitive enzyme immunoassay process utilizing horseradish peroxidase (HRP)-conjugated SARS-CoV-2 as a reporter binding molecule to compete binding with antibody against the SARS-CoV-2 receptor binding domain (SARS-CoV-2 RBD) protein. This strategy enhanced the current signal via the enzymatic reaction of HRP-conjugated SARS-CoV-2 RBD antibody on the electrode surface. The modification, immobilization, blocking, and detection processes were optimized and evaluated by amperometry. The quantitative analysis of SARS-CoV-2 was conducted based on competitive enzyme immunoassay with amperometric detection using a 3D-printed portable potentiostat for point-of-care COVID-19 diagnosis. The current measurements at -0.2 V yielded a calibration curve with a linear range of 0.01-1500 ng mL-1 (r2 = 0.983), a low detection limit of 2 pg mL-1 and a low quantification limit of 10 pg mL-1. In addition, the analyzed results of practical samples using the developed method were successfully verified with ELISA and RT-PCR. Therefore, the proposed portable electrochemical immunosensor is highly sensitive, rapid, and reliable. Thus, it is an alternative ready-to-use sensor for COVID-19 point-of-care diagnosis.