Quantum dots assembly enhanced and dual-antigen sandwich structured lateral flow immunoassay of SARS-CoV-2 antibody with simultaneously high sensitivity and specificity

Jianghua Jia; Lijiao Ao; Yongxin Luo; Tao Liao; Liang Huang; Dinglv Zhuo; Chenxing Jiang; Jing Wang; Jun Hu

doi:10.1016/j.bios.2021.113810

Quantum dots assembly enhanced and dual-antigen sandwich structured lateral flow immunoassay of SARS-CoV-2 antibody with simultaneously high sensitivity and specificity

Biosens Bioelectron. 2022 Feb 15:198:113810. doi: 10.1016/j.bios.2021.113810. Epub 2021 Nov 17.

Authors

Jianghua Jia¹, Lijiao Ao², Yongxin Luo³, Tao Liao⁴, Liang Huang⁵, Dinglv Zhuo⁶, Chenxing Jiang⁶, Jing Wang⁷, Jun Hu⁸

Affiliations

¹ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, 310014, China; Medicalsystem Biotechnology Co., Ltd, Ningbo, 315104, China.
² Institute of Biomedical Engineering, The Second Clinical Medical College (Shenzhen People's Hospital) of Jinan University, Shenzhen, 518020, China.
³ Xinyu People's Hospital, Xinyu, 338000, China.
⁴ Shenzhen WWHS Biotech. Inc, Shenzhen, 518100, China.
⁵ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: lhuang@zjut.edu.cn.
⁶ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, 310014, China.
⁷ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: Jingw1986@zjut.edu.cn.
⁸ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: hjzjut@zjut.edu.cn.

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.

MeSH terms

Antibodies, Viral
Biosensing Techniques*
COVID-19*
Humans
Immunoassay
Quantum Dots*
SARS-CoV-2
Sensitivity and Specificity

Substances

Antibodies, Viral