以重组PvpA蛋白作为抗原的鸡毒支原体抗体间接ELISA检测方法的建立

摘要

为建立鸡毒支原体(MG)种特异性检测的间接ELISA检测方法,本研究以原核表达的PvpA蛋白作为包被抗原,初步建立了检测MG抗体的间接ELISA检测方法.特异性试验结果表明,重组PvpA蛋白与鸡新城疫病毒、鸡传染性支气管炎病毒、滑液支原体、大肠杆菌“O”抗原、禽沙门氏茵“O”抗原阳性血清均不发生交叉反应.该方法的批内变异系数小于8％,批间变异系数小于11％,表明具有较好的重复性.采用该检测方法与进口试剂盒对不同省份送检的鸡血清进行检测比较,两者阳性和阴性符合率均达到90％以上.本研究建立的间接ELISA检测方法为MG抗体检测试剂盒的研制奠定了基础.%n this study, an indirect ELISA for detecting antibodies against Mycoplasma gallisepticum was established based on the purified cytadhesin protein PvpA expressed in E. Coli as coating antigen. The indirect ELISA showed no cross reaction with the positive serum of Newcastle disease virus, Infectious bronchitis virus, M. Synoviae, E. Coli (O) and Salmonella (O). The variation coefficient was less than 8% for intro-batch and less than 11% for inter-batch. Concordance of the indirect ELISA relative to commercial Mycoplasma Gallisepticum Antibody Kit was above 90%. The results indicated that the indirect ELISA could be used for the diagnosis and epidemiological surveys of M. Gallisepticum infection.