鸡产蛋下降综合症病毒快速纯化及其多抗血清制备

Rapid Purification of Egg Drop Syndrome-76 Virus and Preparation of Its Polyclonal Antibodies

为研制鸡产蛋下降综合症病毒(EDS-76)检测用标准抗原及标准多抗血清,以EDS-76国际通用毒株AV-127株为抗原,进行了EDS-76病毒抗原纯化及多抗血清制备。研究表明,经氯仿抽提,并经PEG浓缩处理病毒的尿囊液可获得良好的纯化效果,纯化病毒背景干净,血凝效价达19Log2。以此研制出的鸡抗EDS-76多抗血清血凝抑制效价达13Log2,间接免疫荧光效价达1∶2000,Western-blot试验分析表明该多抗血清可以识别两个分子大小在50 kD至85 kD之间的蛋白抗原,HI及IFA试验发现所制备的特异性,多抗血清与12种常见禽病病原(NDV、AIV-A、ALV-J、ALV、GPV、REV、MDV、IBV、CAV、IBDV、TMUV、REOV)均不反应。本研究建立的快速简便的EDS-76病毒纯化方法以及获得的抗EDS-76多抗血清,为进一步研制EDS-76标准抗原以及标准多抗血清提供了有效的方法与材料。

To prepare standard antigen and polyclonal antibodies for detection of chicken egg drop syndrome-76 virus （ EDS-76） , we developed a rapid method of virus purification with international strain AV-127. Our results showed that the purified virus particles could be obtained by chloroform extraction and PEG concentration from allantoic fluid. The HA titer of the purified virus reached to 19Log2. The polyclonal antibodies to EDS-76 generated here showed 13Log2 in HI titer and 1 ∶ 2000 in IFA titer. Western-blot analysis results showed the same pattern to the available commercial sera against EDS-76. Moreover, the generated polyclonal antibodies against EDS-76 did not show cross-reaction with 12 other avian pathogens（ NDV, AIV-A, ALV-J, ALV, GPV, REV, MDV, IBV, CAV, IBDV, TMUV, REOV） . Overall, our rapid viral purification method provides efficient tools and materials in developing standard antigen and antibodies for detection of EDS-76 in future.