| H3N8亚型马流感病毒间接ELISA抗体检测方法建立及应用 |
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| 引用本文： | 郭巍,王英原,王宇,张馨心,赵立平,王立群,相文华.H3N8亚型马流感病毒间接ELISA抗体检测方法建立及应用[J].中国预防兽医学报,2010,32(3). |
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| 作者姓名： | 郭巍 王英原 王宇 张馨心 赵立平 王立群 相文华 |
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| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江,哈尔滨,150001
2. 东北农业大学生命科学学院,黑龙江,哈尔滨,150030 |
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| 基金项目： | 黑龙江省自然科学基金重点项目 |
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| 摘 要： | 为建立马流感血清学ELISA诊断方法,本研究以马流感病毒中国分离株A/马/新疆/07(H3N8)通过SPF鸡胚培养和增殖,收取含病毒尿囊液经蔗糖密度梯度离心纯化后作为ELISA包被抗原,首次在我国建立了检测H3N8亚型马流感抗体的间接ELISA诊断方法。试验的最佳反应条件为:最佳抗原稀释度7μg/mL,封闭液5%脱脂乳,血清稀释度1∶100,二抗稀释度1∶10000,稀释液PBS(pH7.4),血清反应时间1.5h,二抗反应时间1h。通过本方法对555份临床样品进行检测并与血凝抑制(HI)试验检测结果比较,证明本方法特异、敏感,具有良好的稳定性和可重复性,适于马流感的流行病学调查和监测工作。
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| 关 键 词： | 马流感 间接ELISA 抗体检测 |
Establishment and application of indirect ELISA for the detection of antibody for equine influenza virus H3N8 subtype |
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| GUO Wei,WANG Ying-yuan,WANG Yu,ZHANG Xin-xin,ZHAO Li-ping,WANG Li-qun,XIANG Wen-hua.Establishment and application of indirect ELISA for the detection of antibody for equine influenza virus H3N8 subtype[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(3). |
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| Authors: | GUO Wei WANG Ying-yuan WANG Yu ZHANG Xin-xin ZHAO Li-ping WANG Li-qun XIANG Wen-hua |
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| Institution: | GUO Wei1,WANG Ying-yuan2,WANG Yu1,ZHANG Xin-xin2,ZHAO Li-ping1,WANG Li-qun2,XIANG Wen-hua2 (1. Division of Livestock Infectious Diseases,State Key Laboratory Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultrual Sciences,Harbin 150001,China,2. College of life science,Northeast Agriculture University,Harbin 150030,China) |
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| Abstract: | An mdkeet ELISA for the detection of antibody of H3N8 subtype equine influenza virus (EIV) was established using an EIV A/Equine/Xinjiang/07 (H3N8) strain isolated from China as standard antigen.The assay utilized an optimal antigen coating concentration of 7μg/m,5%skim milk,1:100 serum dilution and 1:10 000 dilution for 2~(nd) antibodies.The established protocol was used to detct 555 samples,and compared with hemagglutination inhibition (HI) test.The results showed that this indirect ELISA was specific and sensitive for EIV antibody,which would provide a convenient method in studies on the epidemiology and surveillance of equine influenza. |
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| Keywords: | Equine influenza Indirect ELISA detection of antibody |
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