检测鸭坦布苏病毒NS1蛋白抗体ELISA方法的建立
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- 英文论文题名：Establishment of an ELISA Assay for the Detection of NS1 Protein of Duck Tembusu Virus
- 论文作者：孙敏华 ; 郭建伟 ; 李林林 ; 董嘉文 ; 邝瑞欢 ; 吴玄光 ; 张建峰 ; 胡奇林 ; 张春红
- 英文论文作者：Sun Minhua ; Guo Jianwei ; Li Linlin ; Dong Jiawen ; Kuang Ruihuan ; Wu Xuanguang ; Zhang Jianfeng ; Hu Qilin ; Zhang Chunhong ; Guangdong Open Laboratory of Public Health ; Guangdong Laboratory for Animal Diseases ; Institute of Animal Health ; Guangdong Academy of Agricultural Sciences ; College of Veterinary Medicine ; South China Agricultural University ; Institute of Animal Husbandry and Veterinary Medicine ; Fujian Academy of Agricultural Sciences
- 年：2016
- 作者机构：广东省农业科学院动物卫生研究所广东省兽医公共卫生公共实验室广东省畜禽疫病防治研究重点实验室;华南农业大学兽医学院;福建农业科学院畜牧兽医研究所;
- 论文关键词：鸭坦布苏病毒 ; NS1蛋白 ; ELISA
- 英文论文关键词：Duck Tembusu virus ; NS1 protein ; ELISA
- 会议召开时间：2016-06-14
- 会议录名称：第25届广东省科技进步活动月畜牧兽医学术与科技创新发展大会论文集
- 语种：中文
- 分类号：S858.32
- 学会代码：GDKL
- 会议名称：第25届广东省科技进步活动月畜牧兽医学术与科技创新发展大会
- 会议地点：中国广东广州
- 主办单位：广东省畜牧兽医学会
- 学会名称：广东省科学技术协会科技交流部
- 页数：5
- 文件大小：282k
- 原文格式：O
- 会议级别：地方
摘要
为了检测鸭群中鸭坦布苏病毒的血清学流行情况,本研究在前期成功表达鸭坦布苏病毒NS1蛋白的基础上,建立了检测鸭血清中鸭坦布苏病毒抗体的间接ELISA方法。棋盘法确定了当纯化的NS1蛋白包被量为0.188μg/孔,待检血清1:200稀释,HRP标记的兔抗鸭二抗1:16000倍稀释时反应条件最佳。在最佳反应条件下,阴阳性临界值判定标准为0.467。用建立的间接ELISA方法对番鸭细小病毒、鹅细小病毒、鸭肝炎病毒、鸭疫里默氏杆菌阳性血清进行了检测,均无交叉反应,表明该方法具有良好的特异性。批内和批间重复试验的最大变异系数分别为6%和10%,显示该方法具有很好的稳定性。用建立的ELISA方法检测临床鸭坦布苏病毒感染鸭及攻毒蛋鸭的55份临床血清样品,测得52份为阳性,阳性率为94.5%,对36份阴性鸭群血清样品进行检测,36份为阴性,阴性率为1 00%。该方法为鸭坦布苏病毒的诊断和流行病学监测奠定了基础。
To survillance the serological prevalence of Duck Tembusu Virus antibodies in duck flocks,an indirect ELISA(Enzyme Linked Immunosorbent Assay) was established based on the recombinant NSl protein which successfully expressed previously.When NS 1 protein coated with a dosage of 0.188 microgram per well(μg/well),the positive serum undergone 1:200 dilution,and enzyme labelled antibody diluted to 1:16000,the assay was optimal with a cut-off value of 0.467.The high specificity made the assay reacted with none of the these positive sera,including Muscovy duck parvovirus,Goose parvovirus,Duck hepatitis virus,and Riemerella anatipestifer.The intra- and inter- coefficient of variabilities were 6%and 10%respectively,which suggested the assay was stable.Clinical tests showed that 52 of 55 sera collected from infected or challenged ducks were positive,and 36 sera collected from negative ducks all gave negetive results.In conclusion,this method could be useful for the detection and survillence of duck Tembusu virus.
To survillance the serological prevalence of Duck Tembusu Virus antibodies in duck flocks,an indirect ELISA(Enzyme Linked Immunosorbent Assay) was established based on the recombinant NSl protein which successfully expressed previously.When NS 1 protein coated with a dosage of 0.188 microgram per well(μg/well),the positive serum undergone 1:200 dilution,and enzyme labelled antibody diluted to 1:16000,the assay was optimal with a cut-off value of 0.467.The high specificity made the assay reacted with none of the these positive sera,including Muscovy duck parvovirus,Goose parvovirus,Duck hepatitis virus,and Riemerella anatipestifer.The intra- and inter- coefficient of variabilities were 6%and 10%respectively,which suggested the assay was stable.Clinical tests showed that 52 of 55 sera collected from infected or challenged ducks were positive,and 36 sera collected from negative ducks all gave negetive results.In conclusion,this method could be useful for the detection and survillence of duck Tembusu virus.
引文
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