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 Cu2+, which prevented the reduction of Cu2+ 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 H2O2. 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.