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为了研究牛支原体P48抗原蛋白的免疫学功能及相应卵黄抗体的生物学特性,探究其抗体效价随免疫时间的变化规律,试验采用牛支原体P48蛋白疫苗免疫健康产蛋母鸡,定时采血并收集鸡蛋,用水稀释法结合硫酸铵盐析法制备特异性卵黄抗体,利用间接ELISA法检测血清抗体(IgG)和卵黄抗体(IgY)效价。结果表明,低、中、高剂量组均能诱导蛋鸡产生有效免疫应答,血清抗体效价与卵黄抗体效价的变化趋势基本一致,但卵黄抗体的产生滞后于血清抗体,在首次免疫后8~10周Ig Y效价达到峰值,其中中剂量组的免疫效价最高,最高效价为1:212。经SDS-PAGE检测,制备所得的纯度达86%以上,得率在8.6 mg/m L以上。说明采用牛支原体P48蛋白免疫蛋鸡可制备高效价、高纯度的抗牛支原体P48蛋白卵黄抗体,从而为卵黄抗体在牛支原体的检测与防控方面的研究奠定了基础。
In order to study the immunological function of P48 antigen protein of M. bovis and the biological characteristics of the corresponding yolk antibody, the change trend of its antibody titer with the immunization time was explored. The healthy pregnant laying hens were immunized with the P48 protein vaccine of M. bovis, Eggs, specific yolk antibodies were prepared by dilution with water and ammonium sulfate, and the serum antibody (IgG) and yolk antibody (IgY) titers were detected by indirect ELISA. The results showed that low, medium and high dose group can induce effective immune response in laying hens, the trend of serum antibody titer and vitellin antibody titer are basically the same, but the yolk antibody lags behind the serum antibody. After the first immunization 8 ~ 10 weeks Ig Y titer reached its peak, of which the highest dose of the highest immune titer, the highest titer of 1: 212. The purity of the obtained product was over 86% by SDS-PAGE and the yield was above 8.6 mg / m L. The results showed that the preparation of P48 protein yolk antibody against Mycoplasma bovine mycoplasma by immunizing hens with P48 protein of M. bovis lays the foundation for the detection of Yolk antibodies in the detection and prevention and control of M. bovis.