摘要
为建立检测流产嗜性衣原体(C.abortus)抗体的快速检测方法,本研究以纯化的重组表达C.abortus主要外膜蛋白(MOMP)为包被抗原,采用方阵法确定包被抗原和待检血清的最佳工作浓度,对反应条件进行优化,建立了C.abortus抗体的间接ELISA检测方法。该方法与猪流行性乙型脑炎病毒、猪瘟病毒、猪伪狂犬病毒、猪繁殖与呼吸障碍综合征病毒阳性血清均无交叉反应;该方法检测稀释800倍的C.abortus阳性血清仍呈阳性;批内和批间变异系数均小于10%。采用所建立的ELISA方法与间接血凝(IHA)方法分别对62份临床血清样品进行检测,阳性率分别为83.87%(52/62)和70.97%(44/62),符合率为87.09%(54/62)。由此表明,本实验建立的间接ELISA方法具有较高的敏感性和特异性,适于C.abortus流行病学调查。
In this study, an indirect ELISA （MOMP-iELISA） for detecting antibodies against Chlamydophila abortu was developed by using recombinant major outer membrane protein （MOMP） as the coating antigen. According the results of specific assay and repetitive testing, this MOMP-iELISA was specific for C.abortu detection, and had no cross reaction with positive serum of Japanese encephalitis virus, classic swine fever virus, porcine pseudorabies virus and porcine reproductie and respiratory syndrome virus. Both intra- and inter-batch coefficient of variation were less than 10%. A total of 62 random serum samples from 5 large scale pig farms were detected by both MOMP-iELISA and IHA, and the positive rate were 83.87% （52/62） and 70.97% （44/62）, respectively, and coincidence rate was 87.09% （54/62）. These results demonstrated that MOMP-iELISA was more sensitive than IHA which was widely used in clinical detection for C.abortu infection.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2015年第9期703-706,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
湖北省重点新产品新工艺研究开发项目(2012BBA15009)
湖北省科技支撑计划(2013BEC0002)
湖北省农业科技创新中心项目(2011-620-001-003)
湖北省自然科学基金(2012FFB01404)
