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目的评价CV-A16-K168免疫血清对CV-A16不同毒株的中和抗体效价,确定完全中和不同病毒量所需的最低血清量,为CV-A16疫苗毒种检定中的鉴别试验和病毒外源因子检查提供参考。方法将CVA16病毒抗原辅以弗氏佐剂以皮下多点注射方式免疫新西兰大白兔,多次免疫后取血清,采用微量细胞病变法进行中和抗体效价检测和对15株不同CV-A16毒株的中和效价检测,并用CV-A16-4V和CV-A16-10R两个毒株分析中和不同量病毒所需的抗体量。结果浓缩抗原作为免疫原获得的兔抗CV-A16血清中和抗体效价可达1∶2 048;该免疫血清对15株CV-A16不同毒株的几何平均滴度(genomic mean titer,GMT)为1∶3 118;完全中和7.5lgCCID50/ml CV-A16病毒所需的最低血清量为128U,当病毒量在4.0~7.0lgCCID50/ml时,完全中和所需的最低血清量为48U,当病毒量低于4.0lgCCID50/ml时,1U的血清即可完全中和本病毒。结论 CV-A16-K168株作为免疫原可获得高效价的抗血清;用不同毒株检测得到的抗体效价存在差异;完全中和病毒所需的最低血清量与病毒量呈正相关;病毒量与测得的血清中和效价呈负相关。
Objective To evaluate the neutralizing antibody titer of CV-A16-K168 immune sera against different strains of CV-A16 and to determine the minimum serum level needed to completely neutralize different viral loads. Virus exogenous factor examination for reference. Methods New Zealand white rabbits were immunized with multiple injections of CVA16 antigen and Freund’s adjuvant. After multiple immunization, serum was taken and the titer of neutralizing antibody was detected by micro-cytopathic assay. The strains were tested for neutralization titer and the amount of antibody required to neutralize different amounts of virus was analyzed using both CV-A16-4V and CV-A16-10R strains. Results The titer of the rabbit anti-CV-A16 serum neutralizing antibody obtained as antigen was 1: 2 048. The immunological serum showed significant differences in the mean genomic mean titer (GMT) of 15 different strains of CV-A16, Is 1: 3 118; the minimum serum requirement for complete neutralization of 7.5 lg CCID50 / ml CV-A16 virus is 128 U, the minimum serum requirement for complete neutralization is 48 U when the viral load is 4.0-7.0 lg CCID50 / ml, When the amount of virus is less than 4.0lgCCID50 / ml, 1U of serum can completely neutralize the virus. Conclusions CV-A16-K168 strain can be used as immunogen to obtain antiserum with high titer. The titer of antibodies detected by different strains is different. The minimum amount of serum required for complete neutralization of virus is positively correlated with the amount of virus. The measured serum neutralization titer was negatively correlated.