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Chemiluminescent Imaging Assay of SARS‐CoV‐2 Protein with Target‐Induced Enzyme Activity Regulation
Author(s) -
Chen Yuhui,
Ao Hang,
Xiao Wencheng,
Ju Huangxian,
Wu Jie
Publication year - 2022
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.202201425
Subject(s) - chemiluminescence , horseradish peroxidase , luminol , chemistry , covid-19 , pyrophosphate , enzyme , coronavirus , microbiology and biotechnology , virology , peroxidase , molecular beacon , enzyme assay , biochemistry , chromatography , dna , biology , medicine , pathology , oligonucleotide , disease , infectious disease (medical specialty)
Simple but robust testing assays are essential for screening and diagnosis of individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) in COVID‐19 pandemic. Here, we described a chemiluminescent imaging assay (CLIA) for sensitive and convenient detection of SARS‐CoV‐2 nucleocapsid protein (NP) by a target‐induced enzyme activity regulation (T‐EAR) strategy. The T‐EAR used a pair of antibody‐DNA probes to recognize SARS‐CoV‐2 NP and proximity‐induce rolling circle amplification for mass‐production of pyrophosphate to coordinate with Cu 2+ , which prevented the reduction of Cu 2+ to Cu + by sodium ascorbate as well as the Cu + ‐caused inactivation of horseradish peroxidase (HRP). The activity retention of HRP produced strong CL signal for the detection of SARS‐CoV‐2 NP by catalyzing the oxidation of luminol by H 2 O 2 . The T‐EAR based CLIA showed a wide detection range from 1 pg/mL to 100 ng/mL (13 fM to 1.3 nM) with the requirement of only 0.75 μL of sample. This CLIA had advantages of good sensitivity, simple wash‐free operation, acceptable accuracy, and high‐throughput imaging detection, displaying potential applicability in screening assay of COVID‐19 infection.