目的  制备抗脊髓灰质炎（脊灰）病毒血清作为检测参考品，并研究其稳定性。方法  用1、2、3型脊灰病毒分别接种非洲绿猴肾细胞，收获病毒液，超滤浓缩后纯化病毒抗原，并进行蛋白含量和纯度检测。用纯化的病毒抗原免疫新西兰大白兔获得分别抗3个型别的血清，过滤、分装、冻干后，进行无菌检验、支原体检验、水分测定，并通过观察细胞病变抑制法进行特异性检测。3名实验员各标定5次候选参考品的中和抗体效价，计算变异系数。血清放置于-20 ℃进行稳定性分析。结果  纯化的1、2、3型脊灰病毒抗原蛋白含量分别为12.3、10.4、9.8 μg/ml。电泳检测中蛋白条带的相对分子质量与文献报道一致。高效液相色谱法检测的1、2、3型蛋白纯度分别为100%、99%和96%。抗各型脊灰病毒血清仅可中和对应型别，而不能中和另两个型别脊灰病毒或肠道病毒。无菌检验、支原体检验结果及水分含量（均低于3%）均符合药典要求。血清参考品的抗1、2、3型脊灰病毒中和抗体效价分别为73 587 、3 264、34 857。候选参考品在-20 ℃放置5年后中和抗体效价未降低（t=-1.25，P=0.225）。结论  制备的抗1、2、3型脊灰病毒冻干血清参考品稳定性好，适用于检测Sabin株脊灰灭活疫苗免疫原性及脊灰病毒特异性中和抗体。

Objective  To prepare anti-poliovirus reference serum and study its stability. Methods  Polioviruses type 1/2/3 were cultured in African Green Monkey Kidney cells, harvested, concentrated by ultrafiltration, and purified by chromatography. The protein contents and purity of virus antigens were analyzed. Anti-sera were obtained from New Zealand white rabbits injected with purified virus antigens, filtered, aliquoted, and lyophilized, before subjected to sterility and mycoplasma tests, and moisture measurement. The specificity was determined by microcytopathic effect inhibition assay. Neutralizing antibody titers of candidate reference sera were calibrated by three experimenters and coefficients of variation were calculated. Sera were stored at -20 ℃ for stability test. Results  The protein contents of purified poliovirus type 1/2/3 bulk were 12.3, 10.4, 9.8 μg /ml, respectively. The relative molecular masses of protein bands seen on electrophoresis gel corresponded to previous report. Protein purities detected by high performance liquid chromatography were 100%, 99%, 96%, respectively. Each type of serum could only neutralize the specific poliovirus type, but not the other two types or enteroviruses. The sterility and mycoplasmas results of candidate in-house reference sera and residual moisture contents (all less than 3%), were in line with quality requirements in pharmacopoeia. The neutralizing antibody titers were 73 587, 3 264, and 34 857 for polioviruses type 1/2/3, respectively. Titer maintained stable for five years at -20 ℃. Conclusion  The in-house reference sera prepared for poliovirus have high stability and are suitable for the detection of Sabin strain inactivated poliovirus vaccine immunogenicity and specific neutralization antibodies against polivirus.