鹿流行性出血病病毒双抗夹心ELISA方法的建立  

Establishment of Sandwich ELISA for Epizootic Hemorrhagic Disease of Deer Virus

作　　者：陈兵[1] 李健波[1] 杨俊兴[1] 阮周曦[1] 孙洁[1] 张彩虹[1] 刘建利[1] 花群义[1] 周晓黎[2] 

机构地区：[1]深圳出入境检验检疫局,广东深圳518001 [2]云南出入境检验检疫局,云南昆明650228

出　　处：《动物医学进展》2011年第10期1-5,共5页Progress In Veterinary Medicine

基　　金：国家863计划项目(2006AA10Z445);中国博士后科学基金(20080430855)

摘　　要：为建立鹿流行性出血病病毒(EHDV)病原学检测方法,用纯化的抗EHDV特异性单克隆抗体包被ELISA板,用兔抗EHDV IgG作为夹心抗体,IgG作为夹心抗体建立EHDV双抗夹心ELISA方法,并对该方法的特异性和敏感性进行了试验。用ELISA分别检测EHDV、蓝舌病病毒(BTV)、水疱性口炎病毒(VSV)、赤羽病病毒(AKV)、小反刍兽疫病毒(PPRV)样品,用RT-PCR作对照。结果表明,ELISA具有良好的特异性和敏感性。用夹心ELISA和RT-PCR同时检测128份参考样品,结果表明夹心ELISA的特异性和敏感性分别为100%和96.3%,两种方法的符合率为99.2%。本研究结果为EHDV检测及鹿流行性出血病流行病学调查提供了有效的工具。In order to establish an immunoassay for Epizootic hemorrhagic disease of deer virus（EHDV） detection,a double-antibody sandwich ELISA was developed based on purified monoclonal antibody against EHDV and rabbit anti EHDV polyclonal antibody.This ELISA was used to detect positive samples of EHDV,bluetongue virus（BTV）,Vesicular stomatitis virus（VSV）,Akabane disease virus（AKV） and Peste des petits ruminants virus（PPRV） and negative samples,and the results indicated that the ELISA had good specificity.This ELISA was also used to test 128 reference samples,RT-PCR was used as a reference method,the results showed the specificity and sensitivity were 100% and 96.3% respectively,the agreement ratio between ELISA and RT-PCR was 99.2%.This ELISA is suitable for rapid detection of EHDV infection in ruminants.

关 键 词：鹿流行性出病 鹿流行性出病病毒 双抗体夹心ELISA 单克隆抗体 

分 类 号：S854.43[农业科学—临床兽医学]