Abstract: Objective To develope a method for the preparation of pseudotype vesicular stomatitis virus(VSV) with chikungunya virus(CHIKV)envelop protein.Methods VSV without G protein genome was used as the vector to produce pseudotype VSV.Results Pseudotype VSV with CHIKV envelop protein and green fluorescent protein(GFP) was developed.In the efficacy test,the infectivity of the pseudotype VSV constructed was 2.73% for the 1:2000 dilution of the antibody against CHIKV E2 of VSV-E2E1 virus,9.61% for the dilution of 1:4000,19.76% for the dilution of 1:8000,29.23% for the dilution of 1:16000,41.68% for the dilution of 1:32000,and 74.84% for the dilution of 1:64000,respectively;the infectivity of negative control was 2.46%.The viral replication was mostly inhibited under the dilution of 1:2000 for the antibody against CHIKV E2.Conclusion The constructed pseudotype VSV is safe and stable and can be applied in epidemic survey by measuring the expression of GFP in infected cells or the activity of luciferase.