单向免疫扩散测定四价流感病毒裂解疫苗中两种B型血凝素含量

Determination of the contents of two types of hemagglutinin B in quadrivalent influenza vaccines (split virion) by single radial immunodiffusion assay

目的四价流感病毒裂解疫苗在三价疫苗中新增一个谱系的B型血凝素抗原,单向免疫扩散法(singleradial immunodiffusion,SRID)是否能准确测定四价疫苗中两种谱系的B型血凝素抗原含量。方法采用江苏金迪克生物技术有限公司研制的四价流感病毒裂解疫苗,运用SRID进行测定。结果在正常检测范围内(10~40μg/ml)比较标准品抗原对照、单价样品、B1+B2单价样品的血凝素测定值差异无统计学意义(P>0.05)。在10~160μg/ml检测范围内分别以三价苗和固定浓度30μg/ml的B型血凝素为基质稀释待测的另一B型血凝素,与该B型不同稀释度的单一血凝素测定结果进行比较,以测定两种B型流感血凝素有无互相干扰。在流感疫苗血凝素检测范围10~40μg/ml之内时,检测的血凝素含量与沉淀环直径呈正比关系(决定系数R^2=0.998),而在10~160μg/ml检测范围内,三元线性回归方程拟合度最高(决定系数R^2=0.999)。结论单向免疫扩散法检测B1和B2毒株的血凝素抗原在10~40μg/ml和10~160μg/ml范围内和存在其他三种血凝素或另一B型血凝素时,均未见互相干扰。

Objective Quadrivalent influenza vaccines contain two lineages of type B virus, this study aimed to assess whether the result of single radial immunodiffusion (SRID) are accurate. The cross-interference of two type B hemagglutinins remains unknown. Methods We detected the vaccine samples developed by Jiangsu GDK Biological Technology Co., ltd by SRID. Results There was no significant difference between the HA content of antigen reagent, bulk sample and mixed sample of two B bulk within 10 to 40 μg/ml (P>0.05). Then each hemagglutinin B was diluted respectively by other three HA, 30 μg/ml, the other hemagglutinin B or phosphate buffer solution were measured within 10-160 μg/ml. Within 10-40 μg/ml, the HA content was proportional to the diameters of immunodiffusion (R^2=0.998), while within the higher content range, a ternary linear regression equation fitted best (R^2=0.999). Conclusions No cross-interference between B/Brisbane and B/Phuket was found in SRID within both detection ranges.