Fluorescence-Quenching Lateral Flow Immunoassay for "Turn-On" and Sensitive Detection of Anti-SARS-Cov-2 Neutralizing Antibodies in Human Serum

The titer of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibodies (NAbs) in the human body is an essential reference for evaluating the acquired protective immunity and resistance to SARS-CoV-2 infection. In this study, a fluorescence-quenching lateral flow immunoassay (FQ-LFIA) is established for quantitative detection of anti-SARS-CoV-2 NAbs in the sera of individuals who are vaccinated or infected within 10 min. The ultrabright aggregation-induced emission properties encapsulated in nanoparticles, AIE(490)NP, are applied in the established FQ-LFIA with gold nanoparticles to achieve a fluorescence "turn-on" competitive immunoassay. Under optimized conditions, the FQ-LFIA quantitatively detected 103 positive and 50 negative human serum samples with a limit of detection (LoD) of 1.29 IU mL(-1). A strong correlation is present with the conventional pseudovirus-based virus neutralization test (R-2 = 0.9796, P < 0.0001). In contrast, the traditional LFIA with a "turn-off" mode can only achieve a LoD of 11.06 IU mL(-1). The FQ-LFIA showed excellent sensitivity to anti-SARS-CoV-2 NAbs. The intra- and inter-assay precisions of the established method are below 15%. The established FQ-LFIA has promising potential as a rapid and quantitative method for detecting anti-SARS-CoV-2 NAbs. FQ-LFIA can also be used to detect various biomarkers.