| 1型鸭疫里氏杆菌抗体间接ELISA检测方法的建立 |
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| 引用本文： | 杨依霏,罗青平,张蓉蓉,艾地云,廖永洪,邵华斌,程国富.1型鸭疫里氏杆菌抗体间接ELISA检测方法的建立[J].动物医学进展,2012(1):13-18. |
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| 作者姓名： | 杨依霏 罗青平 张蓉蓉 艾地云 廖永洪 邵华斌 程国富 |
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| 作者单位： | 华中农业大学动物医学院动物病理实验室;湖北省农业科学院畜牧兽医研究所 |
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| 基金项目： | 公益性行业(农业)科研专项:水禽主要疫病快速诊断与疫苗研制(201003012);湖北省科技厅十一五重大攻关项目:规模化养鸭疫病防控技术集成研究与示范(2009BBB003) |
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| 摘 要： | 以1型鸭疫里氏杆菌(RA)全基因组保守区域ompA基因的重组表达产物为包被抗原,建立了检测RA血清抗体的间接ELISA方法。原核表达的重组ompA蛋白经纯化后作为包被物,以方阵滴定法确定抗原最佳包被浓度为1.5μg/mL,待检血清最佳稀释度为1∶100。与普通的微量凝集试验相比较,该方法灵敏度高于凝集试验约16倍～128倍。与鸭源大肠埃希菌、鸭源多杀性巴氏杆菌、鸭链球菌、鸭源呼肠病毒、鸭肝炎病毒和鸭瘟病毒感染鸭血清均无交叉反应,表明该方法特异性好。利用该方法检测了雏鸭免疫RA灭活油乳剂疫苗之后血清抗体水平。
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| 关 键 词： | 鸭疫里氏杆菌 重组ompA蛋白 ELISA |
Development and Application of An Indirect ELISA for Detection of Riemerella anatipestifer Serotype 1 |
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| YANG Yi-fei,LUO Qing-ping,ZHANG Rong-rong,AI Di-yun,LIAO Yong-hong,SHAO Hua-bin,CHENG Guo-fu.Development and Application of An Indirect ELISA for Detection of Riemerella anatipestifer Serotype 1[J].Progress In Veterinary Medicine,2012(1):13-18. |
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| Authors: | YANG Yi-fei LUO Qing-ping ZHANG Rong-rong AI Di-yun LIAO Yong-hong SHAO Hua-bin CHENG Guo-fu |
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| Institution: | 1(1.Veternary Pathology Laboratory of Academe of Veterinary Medicine,Huazhong Agricultural University,Wuhan,Hubei,430070,China; 2.Insitude of Animal Husbandry and Veterinary,Hubei Academy of Academy of Agricultural Science,Wuhan,Hubei,430064,China) |
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| Abstract: | In this research,an indirect enzyme-linked immunosorbent assay(ELISA) was developed to facilitate early detection of R.anatipestifer infection in ducks.The antigen used was a recombinant 42 ku(ompA) protein,which was the product of the conserved ompA gene in Riemerella anatipestifer type 1 genome.The ompA-based ELISA successfully detected ompA antibody of R.anatipestifer serotype 1.The coated material was recombinant protein ompA after purified.The concentration of antigen was 1.5 μg/mL.Test serum was used after dilution 1∶100.Compared with common micro-agglutination test,ELISA in this research was more sensitive which was 16-128 times higher.There were no cross-reaction with E.coli,P.multocidam,Steptococcus,reovirus,duck hepatitis virus type Ⅰand duck plague virus from ducks,which mean the method was specific.With this method,we test the varition of antibody after injection of inactivated oil emulsion vaccine of RA. |
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| Keywords: | Riemerella anatipestifer recombinat protein ompA ELISA |
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