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  

Development of an indirect ELISA for detecting antibodies against Mycoplasma gallisepticum based on recombinant PvpA protein

作　　者：周云雷[1] 李健[1] 魏飞龙[1] 张小华[1] 蒋红霞[1] 

机构地区：[1]华南农业大学兽医学院/广东省兽药研制与安全评价重点实验室,广东广州510642

出　　处：《中国预防兽医学报》2011年第9期713-717,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金项目(30871890;U0631006)

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、禽沙门氏菌"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。In 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.

关 键 词：鸡毒支原体 pvpA 原核表达 间接ELISA 

分 类 号：S852.5[农业科学—基础兽医学]