应用单向免疫扩散法测定亚单位流感疫苗血凝素含量的验证.pdfVIP

应用单向免疫扩散法测定亚单位流感疫苗血凝素含量的验证 田文莉，高燕燕，祁骥，陈群，李小强 （天士力金纳生物技术(天津)有限公司 天津 300410 ） 摘要：目的 对应用单向免疫扩散法检测流感病毒亚单位疫苗血凝素含量的方法进行验证。 方法 该方法采用不同浓度的标 准抗原作为定量参考品与待测抗原一同进行单向免疫扩散，根据沉淀圈与抗原浓度的线性关系对待测抗原进行定量。在对该 方法的验证中，分别验证了标准曲线的线性、实验的重复性与重现性、回收率、专属性、耐用性。 结果 经过验证，该方法 标准曲线R2均大于0.98，相同型别标准曲线之间的斜率和截距的变异系数（CV）均小于10%；实验之间与实验人员之间测得的 成品血凝素含量的CV值均小于2%；平均回收率为97%；少量甲醛的存在和反应时间的适当延长均不会对实验结果产生影响。结 论 该方法适用于流感病毒亚单位疫苗血凝素含量的检测。 关键词：单向免疫扩散；流感病毒亚单位疫苗；血凝素；验证 Validation on the Measurement of Hemagglutinin Content of Influenza Subunit Vaccine by Single Radial Immunodiffusion Assay TIAN Wen-li, GAO Yan-yan, QI Ji, CHEN Qun, LI Xiao-qiang (Tasly Jenner(Tianjin) Co., Ltd, Tianjin 300410, China) ABSTRACT: OBJECTIVE To validate on the measurement of haemagglutinin content of influenza subunit vaccine by single radial immunodiffusion. METHODS Standard antigen in a series of HA concentrations were performed in single radial immunodiffusion with samples together. Results were plotted as mean zone diameter versus the antigen concentration on a linear scale. In this research, the linear of the standard curve, repeatability and reproducibility, recovery, specificity, and 2 durability of the method were validated. RESULTS For all the standard curves, R 0.98, CV of the slope ratios and intercepts of same antigen both 10%; CV of HA concentrations between experiments and between personnel both 2%; the mean recovery was 97%; the results were not effected either by the existence of a small amount of formaldehyde or the extension of reaction time. CONCLUSION This method was suitable for the measurement of haemagglutinin content in influenza subunit vaccines. KEY WORDS: Single radial immunodiffusion (SRID); Influenza subunit vaccine; Haemagglutinin(HA); Validation 亚单位疫苗是用致病性病原体的主要免疫原组