Abstract:
In this study, an ELISA kit for rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its surface spike glycoprotein receptor binding domain (RBD) was prepared. The kit consists of a pair of RBD monoclonal antibodies as capture antibodies and detection antibodies which were coupled to horseradish peroxidase (HRP), and determines by square titration to the optimal operating conditions for dual anti-sandwich ELISA. In addition, the linearity, specificity, accuracy and precision of the detection kit were verified. The results showed that the titers of the captured antibody and the microplate detection antibody in the double-anti-sandwich ELISA method were 1:160000 and 1:320000 respectively, and the lower limit of quantitative detection of the kit was 31 ng/mL. Furthmore, the kits have a precision of 0.71%~1.59% and an average accuracy of 96.53%, also no cross-reactivity with other inactivated viruses of the same species. All parameters of the kit meet the 2020 standards of the Chinese Pharmacopoeia (Part Ⅲ), indicating that the kit is suitable for the rapid detection of SARS-CoV-2 and RBD antigen.