目的 建立并验证乙型脑炎减毒活疫苗猪胰蛋白酶残留量ELISA检测方法。方法 应用双抗体夹心ELISA建立乙型脑炎减毒活疫苗猪胰蛋白酶残留量的检测方法，对该方法的准确度、重复性、中间精密度、耐用性、线性、专属性、检测限和定量限进行验证。结果 供试品的加标回收率为92.23%~118.16%。同批供试品的变异系数（coefficient of variation，CV）为6.6%~12.1%。不同实验人员、不同实验日期、不同批次试剂盒检测同批供试品的CV分别为8.1%~11.5%、6.7%~8.5%、8.4%~9.6%。在（37±1） ℃温度范围内供试品回收率差异无统计学意义（F=0.43，P=0.662）。在3.12~200.00 ng/mL范围内线性关系良好，决定系数均大于0.98。方法与供试品基质无交叉反应。检测限和定量限分别为1.25 和3.12 ng/mL。结论 建立的方法准确度、重复性、中间精密度、耐用性、线性、专属性良好，适用于乙型脑炎减毒活疫苗猪胰蛋白酶残留量的检测。

Objective To establish and validate the ELISA method for the determination of porcine trypsin residues in attenuated Japanese encephalitis vaccine (JEV).Methods Double-antibody sandwich ELISA was used to establish a method for the determination of porcine trypsin residues in JEV. The accuracy, repeatability, intermediate precision, durability, linearity, specificity, and detection and quantification limits of the method were verified.Results The recovery rates of test samples ranged from 92.23% to 118.16%. Coefficients of variation (CVs) of the same batch were 6.6%-12.1%. CVs of the same batch was 8.1%-11.5%, 6.7%-8.5%, and 8.4%-9.6%, respectively, for different test personnel, different test dates, and different batch kits. Within the temperature range of （37±1） ℃, the recovery rates of samples showed no statistically significant difference (F=0.43, P=0.662). Coefficients of determination were ＞ 0.98 in the range of 3.12-200.00 ng/mL. The method showed no cross-reaction with the substrate. The limits of detection and quantitation were 1.25 ng/mL and 3.12 ng/mL, respectively.Conclusion The method has good accuracy, repeatability, intermediate precision, durability, linearity and specificity, thus is suitable for the determination of porcine trypsin residues in JEV.