Abstract
Exploring reliable and highly-sensitive SARS-CoV-2 antibody diagnosis by point-of-care (POC) manner, holds great public health significance for extensive COVID-19 screening and controlling. Unfortunately, the currently applied gold based lateral flow immunoassay (GLFIA) may expose both false-negative and false-positive interpretations owing to the sensitivity and specificity limitations, which may cause significant risk and waste of public resources for large population screening. To simultaneously overcome the drawbacks of GLFIA, a novel fluorescent LFIA based on signal amplification and dual-antigen sandwich structure was established with largely improved sensitivity and specificity. The compact three-dimensional incorporation of hydrophobic quantum dots within dendritic affinity templates and multilayer surface derivation guaranteed a high and robust fluorescence of single label, which lowered the false negative rate of GLFIA prominently. A dual-antigen sandwich structure using labeled/immobilized SARS-CoV-2 spike receptor binding domain antigen for capturing total human SARS-CoV-2 antibody was developed, instead of general indirect antibody capturing approach, to reduce the false positive rate of GLFIA. Over 300 cases of COVID-19 negative and 97 cases of COVID-19 positive samples, the current assay revealed a 100% sensitivity and 100% specificity confirmed by both polymerase chain reaction (PCR) and chemiluminescence immunoassay (CLIA), compared with the considerable misinterpretation cases by currently applied GLFIA. The quantitative results verified by receiver operating characteristic curve and other statistical analysis indicated a well-distinguished positive/negative sample groups. The proposed strategy is highly sensitive towards low concentrated SARS-CoV-2 antibody serums and highly specific towards serums from COVID-19 negative persons and patients infected by other viruses.
Keywords: Dual-antigen sandwich immunoassay; Misinterpretation; Point-of-care testing; Quantum dots assembly; Signal amplification.
【저자키워드】 signal amplification, point-of-care testing, Dual-antigen sandwich immunoassay, Misinterpretation, Quantum dots assembly, 【초록키워드】 COVID-19, viruses, public health, CLIA, antibody, POC, Population screening, Diagnosis, risk, Receptor binding domain, Antigen, point-of-care, amplification, lateral flow immunoassay, serum, sensitivity, specificity, PCR, SARS-CoV-2 antibody, Sensitivity and specificity, False negative rate, chemiluminescence immunoassay, Patient, Interpretation, LFIA, characteristic, resource, Quantitative, false-negative, False-positive, Quantum dot, SARS-CoV-2 spike, statistical analysis, signal, hydrophobic, fluorescent, dendritic, positive, human SARS-CoV-2, over, limitations, approach, false positive rate, polymerase chain, robust, indicated, applied, overcome, reduce, groups, COVID-19 negative, 【제목키워드】 lateral flow immunoassay, SARS-CoV-2 antibody, Sensitivity and specificity, Quantum dot,