中文摘要：

以鹅源坦布苏病毒JS804毒株制备的多克隆抗体为捕获抗体,抗鹅坦布苏病毒NS1蛋白的单克隆抗体4A9作为检测抗体,建立了检测坦布苏病毒的双抗体夹心ELISA方法。该方法的最佳反应条件为：兔抗鹅坦布苏病毒多克隆抗体的最适稀释度为1∶1 600,单克隆抗体的最适稀释度为1∶160,样品反应时间为1 h,酶标羊抗鼠二抗工作浓度为1∶5 000,以OD450nm≥0.245作为阳性判定标准。该方法的板间、板内重复性变异系数小于10%,病毒最低检测量为386.25TCID50/0.1 mL,与新城疫病毒（NDV）、禽流感病毒（AIV）、传染性支气管炎病毒（IBV）、鸭肝炎病毒（DHV）和鹅细小病毒（GPV）无交叉反应。应用该方法与RT-PCR方法同时检测22份临床样品,符合率达到86.36%。结果表明,建立的双抗体夹心ELISA方法具有特异性好、灵敏性高和重复性好等优点,可用于坦布苏病毒的快速检测。

英文摘要：

In order to establish double antibody sandwich enzyme-linked immunosorbent assay （DAS-ELISA） for detection of Tembusu virus in geese, polyclonal antibody against Tembusu virus strain JS804 in geese and monoclonal antibody against NS1 protein of the Flavivirus strain were used as the capture antibody and detecting antibody, respectively. The optimal dilutions of the two antibodies were 1 ： 1600 and 1 ： 160 by check-board titration, respectively. The reaction time of sample was 1 hour, and the optimal working dilution of HRP-labelled goat-anti-mouse lgG was 1 ： 5000. The positive standard value was 0. 245 （ OD450nm ）. The variation coefficient of reproducibility was less than 10% , and at least 386.25TCID50/0. 1 mL antigen could be detectable. The ELISA had no cross-reaction with Newcastle disease virus （NDV）, avian influenza virus （AIV） , infectious bronchitis virus （IBV）, duck hepatitis virus （DHV）, and gosling plague virus （GPV）. Twenty-two clinical samples were detected by the ELISA and RT-PCR and the agreement rate was up to 86. 36%. The results revealed that the established ELISA in this study possessed good specificity and reproducibility, and higher sensitivity, indicating a suitable method for rapid detection of Tembusu virus in geese.