间接ELISA检测奶牛乳房炎多联苗抗体方法的研究

Establishment of an indirect ELISA method for detecting serum antibody of inactived multi-vaccine against cattle mastitis

利用超声波破碎法制备无乳链球菌、停乳链球菌和金黄色葡萄球菌可溶性抗原,对3种破碎抗原最佳包被浓度及包被条件、血清样品工作浓度、封闭液、酶标二抗的最佳工作浓度及作用时间等反应体系进行筛选和优化,初步建立了间接ELISA法检测奶牛乳房炎多联苗3种菌血清抗体的方法。对24头泌乳牛进行抗体检测,结果表明该3种抗原反应体系一致,3种抗原最适包被浓度为4-6μg/mL,血清样品最佳稀释度为1∶200,该方法具有快速、简单、特异性和重复性好等优点。

An indirect ELISA method was developed for detecting serum antibody of inactivated multi - vaccine against cattle mastitis. The soluble antigens of Streptococcus agalactiae, Streptococcus dysgalactiaewere and Staphylococcus aureus were made by ultrasonic method, respectively. Select and optimize reaction s sstem of the indirect ELISA by the optimal coating concentration and coating conditions of three antigens, the optimal concentration and reaction time of serum samples, blocking buffer and enzyme sign second -antibody. To test antibody of 24 lactating cows. Results showed that the reaction system of three kinds of antigens was coincided, the optimal coating concentration of antigens were 4 -6 μg/mL, the optimal dilution of serum sample was 1：200. The established indirect ELISA method had advantages of specificity, accuracy, precision and stability.