禽波氏杆菌外膜蛋白的提取及其免疫原性的检测

Study on extraction and antigenicity of the outer-membrane-protein of Bordetella avium

摘要:

为研究禽波氏杆菌OMP的免疫原性,试验采用超声波破碎、TritonX-100处理技术提取了禽波氏杆菌OMP,采用Bradford方法测定禽波氏杆菌OMP含量,进行了SDS-PAGE检测,然后制备油乳剂OMP免疫抗原,对1日龄雏鸡分别以0.3mL(OMP90μg)、0.5mL(OMP150μg)、0.8mL(OMP240μg)的剂量颈部皮下接种。结果:禽波氏杆菌OMP含量为300μg/mL;禽波氏杆菌OMP最佳免疫剂量为0.5mL/只;通过免疫抗体与攻毒保护相关性测试,抗体效价在1:28以上能抵抗致死量禽波氏杆菌的攻击。据间接ELISA法检测的抗体水平可知,抗体的持续时间足以保护雏鸡避过易感日龄,试验发现OMP具有良好的免疫原性。本试验结果将为禽波氏杆菌OMP单克隆抗体的制备、快速诊断试剂盒的研制、亚单位疫苗的开发奠定良好的基础。

Abstract:

To study the immunogenicity of Bordetella avium OMP, ultrasonic dispersion、TritonX-100 technique were used to extract Bordetella avium OMP. Its content was determined by Bradford method and it was detected by SDS-PAGE. Then OMP immunizing antigen was prepared and 1-day chicken was vaccinated by hypodermic inoculation, with the immunizing does of 0.3mL(OMP90μg), 0.5mL(OMP150μg), 0.8mL(OMP240μg), respectively. Results showed that content of Bordetella avium OMP is 300 g/mL, the best immunizing does is 0.5mL each one and Chickens can be protected against Bordetella avium at fetal dose if antibody titer is over 1∶2⁸. We can see from the antibody level detected by indirect ELISA that antibody can last long enough to help chickens pass susceptible period, so OMP has good immunogenicity. Results of this study lay good foundation for the development of monoclonal antibody to OMP, rapid diagnosis kit and subunit vaccine.