P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mice were immunized with prokaryotical y expressed P65 recombinant protein to prepare monoclonal anti-body. After screening with Mhp whole-cel protein and P65 protein, a specific hy-bridoma cel line, 3G12, was obtained by ELISA. Identification results indicated that the antibody secreted by 3G12 hybridoma cel s could react with P65 protein and Mhp whole-cel protein. According to indirect ELISA assay, 3G12 cel culture super-natant possessed a titer of 1∶12 800 against P65 protein and 1∶3 200 against Mhp whole-cel protein; 3G12 ascites possessed a titer of above 1∶4 000 000 against P65 protein and above 1∶20 000 against Mhp 168 whole-cel protein. After long-term in vitro culture and continuous freezing-thawing, 3G12 cel line could stably secrete antibodies. A monoclonal antibody against P65 protein and Mhp whole-cel protein was successful y obtained in the present study, which provided basis for further in-vestigating the pathogenic mechanism of Mhp and establishing diagnostic methods of Mycoplasmal pneumonia of swine （MPS）.
猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)是引起猪支原体肺炎(Mycoplasma pneumoniae of Swine,MPS)的病原,给养猪业造成巨大的经济损失。由于对具有良好免疫原性的Mhp特异性蛋白缺少深入研究,使得Mhp在致病机制、血清学检测方法和新型疫苗的研究发展上受到一定限制,从而影响了对MPS的控制。本研究综合Mhp主要粘附因子、膜蛋白及细胞质蛋白等主要抗原蛋白的研究进展,为该病原致病机理研究、建立特异性诊断方法和推动基因工程疫苗发展提供一定的理论基础,对MPS的正确诊断、检测、免疫预防和控制也有重要意义。
