猪肺炎支原体P97单抗的生物学特性鉴定及初步应用

对已成功制备的猪肺炎支原体P97原核表达产物单抗进行生物学特性鉴定。鉴定结果为:两株单抗的免疫球蛋白亚类均为IgG1,亲和常数分别为52 mg/L,46 mg/L。单抗腹水中的间接ELISA效价分别为1×106和1×105。两株单抗特异性的识别猪肺炎支原体168株97kD左右的抗原成分。在Dot-ELISA试验中,两株单抗均与猪肺炎支原体反应,而与猪鼻支原体(Mh)、猪絮状支原体(Mf)和鸡毒支原体(Mg)无交叉反应。进一步将该单抗用于单抗阻断ELISA中,检测了36份血清样,其中9份呈阳性,检测结果与IDEXX公司猪肺炎支原体抗体检测试剂盒的结果基本吻合,从而为猪肺炎支原体诊断试剂盒的研制及猪气喘病早期诊断方法的建立奠定了基础。

Identification and Application of Monoclonal Antibody of Mycoplasma Hyopneumoniae P97

In this study,we identified monoclonal antibody of P97 gene of Mycoplasma hyopneumoniae expressed in Escherichia coil,2 strain of monoclonal hybridoma cells A9H7and H11E7were established.Both of them were IgG1 subtype,and their affinity index were 52μg/ml and 46μg/ml.Their ascitic fluids titer of ELISA were 1x106and 1x105 respectively.Two MAbs specifically recognized the antigen of 97KD of Mhp strain 168.In Dot-ELISA,the MAbs could react with Mhp,but neither could bind with M.hyorhinis(Mh),M.flocculare(Mf) and M.gallisepticum(Mg).Furthermore,MAb was used in the monoclonal blocking ELISA for specific detection of antibodies against Mhp.With this method,9/36 serum samples were positive,the same as IDEXX Mhp antibody test kit.Our experi-ment provides the basis for the future researching Mhp test kit and for establishing MPS earlier diagnosis methods.