奶牛新孢子虫重组蛋白NcSRS2t间接ELISA方法的建立及初步应用  

Development and preliminary application of indirect ELISA diagnostic method for Neospora caninum in dairy cattle

作　　者：王春仁[1] 李志莲[2] 翟延庆[1] 倪宏波[1] 陈爱华[1] 陈佳[1] 王宇[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]北京农业职业学院,北京102442

出　　处：《中国预防兽医学报》2009年第2期145-149,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：大庆市科技局攻关项目(SGG2005-032)

摘　　要：以纯化的犬新孢子虫(Neospora caninum)重组蛋白NcSRS2t为抗原包被酶标板,通过对间接ELISA各反应条件的优化,确定了最佳反应条件。采用已建立的间接ELISA反应条件,对218份阴性血清的检测结果进行统计学分析,确定了间接ELISA的判定标准,即OD450nm值大于等于0.32为阳性,小于0.32为阴性。应用建立的间接ELISA方法对128份阴性血清和50份阳性血清(经IDEXX公司新孢子虫抗体检测试剂盒证实)进行检测,结果表明,间接ELISA方法的特异性为93.0%,敏感性为90.0%。采用本试验建立的新孢子虫间接ELISA诊断方法对黑龙江省克山、海林、双城、甘南、克东、富裕、牡丹江、大庆等8个县(市)奶牛场的540份奶牛血清进行了检测。结果表明,被检的540份奶牛血清样本中奶牛新孢子虫的抗体阳性率为13.33%。该间接ELISA诊断方法的建立及初步应用为新孢子虫病诊断试剂盒的研制和新孢子虫病的预防研究奠定了基础。An indirect ELISA were developed using purified Neospora caninum recombinant NcSRS2t antigen. The cut-off value of the assay was determined by statistics analysis based on 218 negative bovine serum samples, with OD equal to or higher than 0.32 being considered positive. Test on fifty positive and 128 negative serum samples from the cow （confirmed by the IDEXX Neospora caninum antibody test kit） showed that the indirect ELISA had 93.0% specificity and 90.0% sensitivity. A Total of 540 sera of dairy cattle obtained from Keshan, Hailin, Shuangcheng Gannan, Kedong, Fuyu, Mudanjiang, Daqing in Heilongjiang province were screened for neosporosis infection by indirect ELISA. The result showed that the seroprevalence of neosporosis in dairy cattle in some regions of Heilongjiang province was 13.33%. This indirect ELISA could be further developed for commercial application in diagnosis and prevention of neosporosis.

关 键 词：新孢子虫 NcSRS2t 间接ELISA 临床应用 

分 类 号：S852.723[农业科学—基础兽医学]