乳胶凝集试验方法在检测鸭坦布苏病毒抗体中的应用
Rapid Detection of Duck Tembusu Virus Antibody by Latex Agglutination Test
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摘要: 以经超速和密度梯度离心浓缩纯化的鸭坦布苏病毒抗原进行方阵滴定, 筛选出致敏乳胶的最佳条件, 并制备成鸭坦布苏病毒乳胶凝集试验 (LAT) 用抗原, 与特异性的鸭坦布苏病毒阳性血清反应出现肉眼可见的凝集颗粒, 进而建立检测鸭坦布苏病毒抗体的乳胶凝集试验方法。经试验测定该致敏的抗原与鸭流感病毒阳性血清、鸭副粘病毒阳性血清、鸭肝炎病毒阳性血清、鸭产蛋综合症病毒阳性血清和鸭呼肠孤病毒阳性血清均不出现凝集现象, 表明具有良好的特异性。以建立的LAT方法和间接ELISA同时分别对80份鸭血清进行检测, 结果 LAT方法检出阳性58份、阴性22份, 而间接ELISA方法检出阳性68份、阴性12份, 两者阳性符合率达85.3%。以上结果表明, 本方法具有简便、快速、特异等优点, 可用于临床样品的快速诊断和血清流行病学调查。Abstract: Duck tembusu viruses were cultivated and concentrated by ultracentrifugation and density-gradient centrifugation to determine the best suitable conditions for sensitized latex and prepare antigen for the latex agglutination test (LAT) by titration.The agglutination particles could be observed clearly by naked eyes after antigen reaction with positive duck tembusu virus serum.The sensitized antigen had no cross-reaction with avian influenza virus, avian paramyxovirus type 1, duck hepatitis virus type 1, egg drop syndrome virus, and duck reovirus.Of 80duck serums, 58and 68 were detected to be positive by the established LAT and ELISA, respectively;the positive rates for both were above 85.3% with no statistical difference.The results revealed that the established LAT was simple, quick and specific for the rapid diagnose and survey of duck tembusu virus antibody.