去垢剂对单向免疫扩散法测定流感病毒疫苗血凝素含量的影响(1)
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[摘要] 目的 研究单向免疫扩散法测定流感病毒亚单位疫苗不同生产阶段中间产品的血凝素含量实验中，去垢剂对检测结果的影响，寻找最佳裂解条件。 方法 以不同的去垢剂浓度和不同裂解时间分别对完整病毒和亚单位疫苗样本进行处理后，利用单向免疫扩散实验测定样本的血凝素含量，并对测定结果进行比较和分析。 结果 亚单位疫苗的单向免疫扩散沉淀圈在去垢剂Zwittergent浓度小于0.125%时随去垢剂浓度的增加而增大，当去垢剂浓度达到0.125%后不再继续增大；而完整病毒的沉淀圈在去垢剂浓度达到1%后才不再增大。以终浓度1%的去垢剂对完整病毒和亚单位疫苗分别处理30、60、120 min后进行单向免疫扩散，测定结果比较差异无统计学意义。 结论 以终浓度1%的去垢剂Zwittergent对完整病毒或亚单位疫苗样本裂解30 min后进行单向免疫扩散测定血凝素含量，均可得到稳定准确的结果，有利于提高工作效率、缩小实验间误差、节约实验成本。
[关键词] 单向免疫扩散；流感病毒疫苗；血凝素；亚单位；去垢剂
[中图分类号] R927.2 [文献标识码] A [文章编号] 1674-4721(2012)02(c)-0049-03
Influence of detergent in the measurement of haemagglutinin content of influenza vaccine by single radial immunodiffusion
TIAN Wenli QI Ji GAO Yanyan CHEN Qun YANG Xu LI Xiaoqiang
Tasly & Jenner (Tianjin) Co., Ltd, Tianjin 300410, China
[Abstract] Objective To research the influence of detergent in single radial immunodiffusion (SRID) test for the assay of haemagglutinin on different influenza vaccine intermedia products, and to determine the disintegration terms treating with detergent. Methods Examined the haemagglutinin content by single radial immunodiffusion assay after treating whole virus and subunit vaccine intermedia products with a variety of concentration and taking time of detergent, compared and analyzed on the measurement data. Results Size of the subunit vaccine diffusion zones increased with increasing concentrations of Zwittergent reaching 0.125%, and kept consistent with detergent concentrations higher than 0.125%. However, size of the whole virus diffusion zones increased until the detergent concentrations reach 1%. The result of haemagglutinin content did not varied by treating with 1% Zwittergent for 30, 60, and 120 min, there were no significant difference among the detergent results. Conclusion Treating with 1% Zwittergent for 30 min is proper for either whole virus or subunit vaccine intermedia products in single radial immunodiffusion for haemagglutinin assay, it is good for increasing the job efficiency, decreasing the experimental error and saving the experimentation costs.
[Key words] Single radial immunodiffusion (SRID); Influenza vaccine; Haemagglutinin(HA); Subunit; Detergent ......
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