Objective To generate monoclonal antibodies (mAbs) against coxsackievirus A2(CV-A2)and to establish a quantitative antigen ELISA. Methods Mice were immunized by purifiedCV-A2 virions to obtain hybridomas secreting anti-CV-A2 mAbs. Linear range of established CV-A2ELISA was determined, and its accuracy, precision, stability and specificity were validated. Samples in all purification steps of CV-A2 particles were detected. Results High binding-efficiency mAbs againstCV-A2 were generated. A quantitative ELISA for antigen detection was established. The detection range was 5. 00-320. 00 ng/ml. The recovery rates for accuracy verification of samples with high, medium andlow concentrations were between 89. 58% and 104. 78%. Their respective coefficients of variation (CVs)of repeatability verification were 2. 10%, 2. 47% and 6. 18%, and CVs of intermediate precision verification were 2. 89%, 2. 69% and 1. 94%. The recovery rates of durability were 84. 26%-114.21%. Coated microtiter plate was placed at 37 ℃ for 3 d and samples recovery rates were90.31%-103.11%. Specificity verification showed that the method only recognized CV-A2 antigen, and did not cross react with other antigens. Conclusion A CV-A2 quantitative ELISA is established, which can be applied to antigen detection in samples at different purification stages of CV-A2, and in multivalent hand-foot-mouth disease vaccines containing CV-A2.