Objective To establish and verify a method for rapid testing of type D antigen in the monovalent bulk of Sabin strain inactivated polio vaccine(sIPV) using surface plasmon resonance(SPR) technology.Methods Bovine polyclonal antibodies against typesⅠ, Ⅱ and Ⅲ D antigens were labeled on the surface of an SPR chip to capture poliovirus typeⅠ, Ⅱ andⅢ D antigens. The change of signal response values on the chip surface was monitored and the relationship between D antigen content and response signals was determined to establish SPR method for the determination of D antigen content. The linearity, specificity, precision and accuracy of the method were validated. The SPR method and ELISA method were used to detect D antigen in six batches of monovalent bulks of sIPV of typeⅠ, Ⅱ and Ⅲ, separately. The results of the two methods were compared and analyzed to supplement the validation of the feasibility of this method.Results The D antigen content of the three types showed a good linear relationship with the response values, with R~2values all above 0. 99. There was some cross-reactivity between the three types, but the cross-reactivity response values were obviously smaller than the positive reaction response values. The relative standard deviations(RSDs) for the repeated measurements of the three types were3. 1%, 1. 2% and 1. 4%, respectively, and the RSDs of intermediate precision results were 5. 6%, 2. 0% and 2. 7%, all within 8%, lower than the 15% limit set by the ELISA method. The recovery rates of accuracy verification were 85. 8%,105. 1% and 88. 4%, all within the range of 80% to 120%. Compared with the ELISA results of six batches of monovalent bulks, there was no statistically significant difference in typeⅠand type Ⅱ(t = 1. 924 and-0. 901, P = 0. 083 and 0. 389,respectively), but a statistically significant difference in type Ⅲ(t =-2. 617, P = 0. 026).Conclusion A SPR method for rapid and high-throughput detection of D antigens in sIPV of typesⅠ, Ⅱ and Ⅲ was established, which has good precision and accuracy, and provides more schemes for the whole process monitoring and quality inspection of vaccine production.