Objective To explore the feasibility of establishing an experimental model of tree shrew lungfibroblasts (TSLF) infected with CA16. Methods TSLF were infected with enterovirus CA16, and human lung fibroblasts(KMB-17) were used as controls. The cytopathic effects ( CPE) were observed using an inverted microscope. Theexpression of viral protein and the SCARB2 receptor protein was observed by indirect immunofluorescence assay, and viralload was detected by TaqMan real-time PCR assay. Using β-actin as an internal reference, the relative expression of theSCARB2 gene was detected using SYBR Green real-time PCR, and its correlation with the infection process was analyzedalong with the gene sequence alignment. Results CA16 infected TSLF and caused obvious CPE. The virus and SCARB2protein were observed by immunofluorescence assay. Under the infection rate of 100TCID₅₀ /10⁵ cells, the viral load washighest 48 h after infection. The SCARB2 gene had a high expression that was associated with infection, and its genesequence also has a high homology with that of humans. Conclusion CA16 can infect TSLF and SCARB2 is related to the infection.