疫苗中残留卵清蛋白ELISA定量测定方法的建立

Quantitative determination of residual ovalbumin in vaccine by ELISA

目的 建立ELISA法检测疫苗中残留卵清蛋白的含量。方法 将进口纯化兔抗卵清蛋白抗体作为包被抗体，辣根过氧化物酶标记兔抗卵清蛋白抗体作为酶标抗体，以系列含量的卵清蛋白溶液为标准，采用ELISA双抗体夹心法，对供试品中残留卵清蛋白进行定量测定。结果 最佳线性范围为1．25～20ng／ml，相关系数r≥0．99；定量限度为1．25ng／ml；准确性试验回收率为89．2％～103．6％；精密性试验内和试验间变异系数分别为5．6％～6．7％和4．2％～9．4％。与马血清、羊血清、人血清白蛋白、牛血清白蛋白、流行性感冒病毒裂解疫苗基质液及其他无残留卵清蛋白的疫苗均无交叉反应。结论 本方法特异性强、灵敏度高，准确性、重复性和稳定性好，可用于疫苗中残留卵清蛋白的定量检测。

Objective To establish a method for determining the content of ovalbumin in vaccines by ELISA. Methods The purified rabbit anti-ovalbumin was used as a coating antibody,a horseradish peroxidase-conjugated anti-ovalbumin as an enzyme-labeled antibody, and a series of concentrations of ovalbumin solution as standards. The content of ovalbumin in samples was determined by a double antibody sandwich ELISA. Results The optimal linear range was 1.25-20 ng/ml and the coefficient of correlation was ≥0.99. The quantitation limit was 1.25 ng/ml. The recovery rate for the accuracy test was 89.2%-103.6%. The coefficients of variation for intra-assay and inter-assay precision were 5.6%-6.7% and 4.2%-9.4% ,respectively. No cross reactions were observed with horse serum, sheep serum,human serum albumin ,bovine serum albumin,diluent for split influenza virus vaccine ,and other vaccines without ovalbumin. Conclusions The method is specific, sensitive, accurate, reproducible,and stable. It is suitable for quantitative determination of residual ovalbumin in vaccines.