鸡传染性法氏囊病毒抗原捕获ELISA检测方法的建立及应用

Development and Application of AC-ELISA for Detection of Infectious Bursal Disease Virus

为快速检测鸡传染性法氏囊病毒,使用纯化的鸡抗IBDV多克隆抗体和鼠抗IBDV单克隆抗体,建立了抗原捕获ELISA检测鸡传染性法氏囊病毒的方法。使用建立的方法可以检测到1∶160倍稀释的IBDV疫苗及阳性法氏囊组织中的病毒。检测禽流感、新城疫、鸡传染性支气管炎病毒及沙门氏菌、葡萄球菌、巴氏杆菌、大肠杆菌、肺炎球菌等结果均为阴性。检测IBDV超强毒和疫苗毒人工感染鸡的法氏囊、脾、肝及临床样品78份,检出阳性24份,与RT-PCR检测结果相符率为96%(75/78)。说明建立的方法敏感、特异,可用于法氏囊病的诊断和监测。

The Antigen Capture Enzyme-Linked Immunosorbent Assay （AC-ELISA） was developed with the purified chicken anti-IBDV antibodies and mouse anti-IBDV VP2 monoclonal antibodies for quickly detecting IBDV. The 1/160 diluted viruses in the IBDV vaccines and the infected bursa of fabricius can be observed, while other pathogens such as AIV, NDV, IBV, Sa/monel/a spp., Staphylococcus aureus, Pasteurella multocida, Escherichia coli and Streptococcus pneumoniea can not be found by the AC-ELISA.78 samples included the bursa of fabricius, spleens, livers which were experimentally infected with very virulent IBDV （wIBDV） and classic viruses in the vaccines, were assayed. The results showed that 24 samples were positive with 96% （75/78） agreement between AC-ELISA and RT-PCR, which indicating that the AC-ELISA with high sensitivity and specificity can be used to diagnose IBD.