Development of receptor binding domain-based double-antigen sandwich lateral flow immunoassay for the detection and evaluation of SARS-CoV-2 neutralizing antibody in clinical sera samples compared with the conventional virus neutralization test.

Vaccination is an effective strategy to fight COVID-19. However, the effectiveness of the vaccine varies among different populations in varying immune effects. Neutralizing antibody (NAb) level is an important indicator to evaluate the protective effect of immune response after vaccination. Lateral flow immunoassay (LFIA) is a rapid, safe and sensitivity detection method, which has great potential in the detection of SARS-CoV-2 NAb. In this study, a fluorescent beads-based lateral flow immunoassay (FBs-LFIA) and a latex beads-based LFIA (LBs-LFIA) using double antigen sandwich (DAS) strategy were established to detect NAbs in the serum of vaccinated people. The limit of detection (LoD) of the FBs-LFIA was 1.13 ng mL− 1 and the LBs-LFIA was 7.11 ng mL− 1. The two LFIAs were no cross-reactive with sera infected by other pathogenic bacteria. Furthermore, the two LFIAs showed a good performance in testing clinical samples. The sensitivity of FBs-LFIA and LBs-LFIA were 97.44% (95%CI: 93.15%–99.18%) and 98.29% (95%CI: 95.84%–99.37%), and the specificity were 98.28% (95%CI: 95.37%–99.45%) and 97.70% (95%CI: 94.82%–99.06%) compared with the conventional virus neutralization test (cVNT), respectively. Notably, the LBs-LFIA was also suitable for whole blood sample, requiring only 3 μL of whole blood, which provided the possibility to detect NAbs at home. To sum up, the two LFIAs based on double antigen sandwich established by us can rapidly, safely, sensitively and accurately detect SARS-CoV-2 NAb in human serum. [Display omitted] • LFIAs based on double antigen sandwich (DAS) have high sensitivity and specificity compared with cVNT. • The FBs-LFIA achieves a sensitivity of 97.44% and specificity of 98.28% in testing clinical serum samples (n = 389). • The LBs-LFIA achieves a sensitivity of 98.29% and a specificity of 97.70% in testing clinical serum samples (n = 554). • The LBs-LFIA can be used to plasma, serum and whole blood samples so it's suitable for home self-test and community hospital. [ABSTRACT FROM AUTHOR]