Chemiluminescent Imaging Assay of SARS-CoV-2 Protein with Target-Induced Enzyme Activity Regulation.
Yuhui ChenHang AoWencheng XiaoHuangxian JuJie WuPublished in: Chemistry (Weinheim an der Bergstrasse, Germany) (2022)
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.
Keyphrases
- sars cov
- respiratory syndrome coronavirus
- high throughput
- high resolution
- label free
- loop mediated isothermal amplification
- real time pcr
- high glucose
- diabetic rats
- small molecule
- fluorescence imaging
- protein protein
- endothelial cells
- aqueous solution
- oxidative stress
- metal organic framework
- nitric oxide
- amino acid
- binding protein
- risk assessment
- cell free
- circulating tumor cells
- stress induced