Objective To develop a standardized ELISA kit for mouse Sendai virus and test it in clinical application. Methods Sendai virus seed batches and BHK-21 cell banks were set firstly; then the crafts for virus production and antigen purification were established; the detection system in ELISA flat was also optimized; the positive and negative serum controls were normalized, a critical concentration positive serum was also set in this system. A totally 672 sera samples were analyzed by the current ELISA kit, and the results were confirmed by IFA and Western blot. Results the Sendai virus seed batches stored at -80℃ were stable for more than 6 months; the virus production craft and antigen purification craft improved the antigen quality; the control systems reduced the effect from environment. The detection sensitivity of ELISA kit is higher than Western blot, much higher than IFA. Conclusion the standardized mouse Sendai virus detection ELISA kit can detect the Sendai virus infection condition, and the detection result is stable and reproducible.