中国莱姆病螺旋体特异性单克隆抗体的制备及初步鉴定

目的制备特异的莱姆病螺旋体单克隆抗体,为我国莱姆病的诊断和莱姆病螺旋体菌株鉴定提供基础。方法以中国莱姆病螺旋体伽氏疏螺旋体(Borreliagarinii)的代表菌株PD91的全菌蛋白为抗原,免疫BALB/c小鼠,取脾细胞与骨髓瘤细胞SP2/0融合,用间接酶联免疫吸附试验(ELISA)和蛋白免疫印迹方法(WB)筛选,并经过2次或3次克隆,以获得单克隆抗体。结果共制备出10株单克隆抗体,经鉴定为3种,分别针对中国莱姆病螺旋体的外膜蛋白OspA(4株)、OspB(3株)和OspC(3株)。结论成功制备出3种抗莱姆病螺旋体不同蛋白的单克隆抗体,可用于我国莱姆病的病原诊断和莱姆病螺旋体菌株鉴定。

Preparation and preliminary identification of monoclonal antibody against Borrelia burgdorferi in China

To prepare and identify the monoclonal antibody against Borrelia burgdorferi in order to provide basis for the detection of Lyme spirochetes and diagnosis of Lyme disease.Spleen cells were prepared from BALB/c mice which was immunized by the whole proteins of B.garinii strain PD91.And then they were fused with SP2/0 myeloma cells.The positive clones were picked out and identified by ELISA and WB.The positive clones were subcloned two or three times to obtain the monoclonal cells.Ten hybridoma cell strains which could produce the monoclonal antibody against B.burgdorferi were obtained,in which four is against OspA,three against OspB and three against OspC.Three kinds of monoclonal antibodies against B.burgdorferi have been successfully prepared.And they can be used in the diagnosis of Lyme disease and detection of Lyme spirochetes.