中文摘要：

为建立快速检测猪流感病毒抗体的血清学方法,利用合成的甲型流感病毒M2蛋白多肽作为包被抗原,建立了检测猪流感病毒血清抗体的间接ELISA,并对各种检测条件进行了优化。结果显示,优化后确定的抗原最适包被量为250ng/孔,血清最佳稀释度为1∶100,酶标二抗最佳稀释度为1∶4 000。在优化条件下,阴阳性临界值的判定标准为0.249。用建立的间接ELISA对CSFV、PRRSV、PCV2、PPV、PRV、FMDV、JEV阳性血清进行了检测,均无交叉反应。用该方法对280份冬季采集的猪血清样品进行检测,有60份样品呈现阳性;同时用IDEXX甲型流感病毒抗体检测试剂盒进行平行检测,有61份样品呈现阳性,两者的符合率达到98.35%。结果表明,建立的ELISA具有良好的敏感性、特异性和可信度。本研究为猪流感的诊断和流行病学调查提供了一种准确、快速、简便的方法。

英文摘要：

An indirect ELISA was established for rapid detection of antibodies against swine influenza virus（SIV） using synthetic M2 protein polypeptide of influenza A virus as coating antigen. The reaction conditions were optimized,including 250 ng/well coating antigen of synthetic SIV M2 protein polypeptide, 1 ： 100 dilution of testing serum and 1 ： 4 000 dilution of HRP-conjugated Rabbit Anti-pig IgG with cut-off value 0. 249（D_450mm）. No cross-reaction was observed with the positive sera against classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine circovirus, porcine parvovirus, pseudo- rabies virus,foot-and-mouth disease virus and Japanese encephalitis virus,respectively. A total of 280 ser- um samples collected in winter were tested,60 samples were positively detected by the indirect ELISA and 61 samples showed positive detected by IDEXX SIV ELISA,the agreement rate was 98.35%. All these re- sults show that the established ELISA method has high sensitivity, reliability and credibility. This study provides an accurate,rapid and simple method for the diagnosis and epidemiological investigation of SIV.