The structural and functional protein VP1 of echovirus is important for echovirus infection.Three antigen epitope-riched fragments of VP1 proteins from three serotype echovirus respectively, type l, 6 and 9, were screened by computer analysis.The gene sequences of the three fragments were optimized,chemically synthesized, and cloned into plasmid pGEX4T-2 respectively for expression and purification by genetic engineering.The antigenicity and immunogenicity of the expressed three fragments were detected by western blotting and immunizing rabbits.The three rabbit polyclonal antibodies to the three VP1 fragments were purified.As the direction antigen, the fragments conjugated with colloidal gold were added onto glass fiber membrane for detecting antibodies against VP1 protein.As test line or control line,the anti-human IgG monoclonal antibody and the anti-VP1 proteins polyclonal antibody were respectively added to the nitrocellulose membranes building the capture reaction system.The colloidal gold test strip was assembled and inspected for its specificity and sensitivity, and was used for detection of antibodies against echovirus in sera of patients.The results showed that the three VP 1 protein fragments from E1, E6 and E9 were efficiently expressed and purified respectively, and could induce strong immune response.The colloidal gold test strip was successfully developed and has good specificity and sensitivity for detection of antibodies against echovirus without cross-reaction with other viruses.