The amino acid sequences of human adenoviruses (HAdV) hexons from five types of HadV, type 3,7, 11, 14 and 55, were analyzed by using Antheprot, and the immunogenic epitopes of each type HAdV hexons were screened, then the DNA sequences of the immunogenic epitopes were optimized, chemically synthesized, linked together, and cloned into plasmid pET-28a(+) for expressing an chimeric protein.The chimeric protein containing the epitopes from five types was expressed and purified by genetic engineering.The purified chimeric protein was synergized with freunds adjuvant for immunizing mice.All of mice were immunized four times.The immunogenicity and antigenicity of chimeric protein were detected by ELISA.Immune colloidal gold technique was used to prepared the rapid detection device.The chimeric protein was linked to colloidal gold to form immune gold.Quality control and detection strings were respectively covered with goat-anti-mouse antibodies and the polyclonal antibodies to the chimeric protein purified from mice sera.The specificity and sensitivity of the colloidal gold device were detected by clinic trial.The results showed that the chimeric protein was efficiently expressed and purified, and immunized successfully.The chimeric protein has strong immunogenicity and antigenicity.The colloidal gold device was prepared successfully, and has good specificity and sensitivity for detecting infection sera of patients.