抗金黄色葡萄球菌卵黄抗体间接ELISA检测方法的建立与应用

Establishment of Indirect ELISA for Detection the level of egg yolk antibody of Staphylococcus aureus Isolated from Co_w

本试验通过矩阵法对最佳包被抗原量及其他ELISA条件进行优化,建立一种检测抗金黄色葡萄球菌卵黄抗体的酶联免疫吸附试验(ELISA)方法,用于免疫后卵黄抗体水平的监测。并将该方法与试管凝集法进行敏感性比较。结果表明,本ELISA法抗原最佳包被量为105个/孔,抗体最佳使用稀释倍数为1:100,酶标兔抗鸡二抗工作浓度为1:2000。通过本试验建立的ELISA方法检测卵黄抗体的水平,发现卵黄抗体在三免后第20天左右达到高峰,阳性水平至少持续到初免后的120d。

Establish an indirect ELISA by phalanx titration to detect the antibody level against staphylococcus aureus after immunization.Meanwhile,the study compared the sensitivity between the indirect ELISA and tube agglutination..The result showed that the optimal concentration of the antigen was 105 piece/cell.The dilutionrate of serum sample was 1：100,the optimal working concentration of the HRP-labeled rabbit anti-chicken IgY was 1：2 000.By the experiment we established the ELISA test which detected the level of egg yolk antibody;we found that egg yolk antibody reached a peak on the 20th day after the third immunization,positive level at least lasting 120 days after the beginning immune.