Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus. Methods The antigen concentration, serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method. The results were compared and the differentia between the two methods was confirmed by IFA. Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1:1, using the concentration of 20 μg/mL and the serum dilution was 1:10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA. Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus. The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps. The method has some advantages in degeneracy and accuracy.