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鸡产蛋下降综合症病毒快速纯化及其抗血清制备
孙苹,秦爱建
0
(扬州大学江苏省动物预防医学重点实验室)
摘要:
为了研制鸡产蛋下降综合症病毒(EDS-76)检测专用标准抗原物质及标准血清物质,本研究以EDS-76标准毒株AV-127株为抗原,进行了EDS-76病毒抗原纯化及抗血清制备。研究发现,简单的氯仿抽提与PEG浓缩处理病毒尿囊液即可获得良好的纯化效果。获得的纯化病毒背景干净,血凝效价达19Log2。以此研制出的鸡抗EDS-76抗血清血凝抑制效价达13Log2,间接免疫荧光效价达1:1000,Western blot试验分析表明该血清可以识别50kDa和70kDa的病毒抗原,且与所检测的其它12种常见禽病病原不反应。综上所述,本研究建立的不依赖于蔗糖梯度超速离心的快速简便的EDS-76病毒纯化方法以及获得的抗EDS-76血清,为进一步研制EDS-76检测专用标准抗原物质以及标准血清物质提供了有效的方法与材料。
关键词:  产蛋下降综合症病毒  纯化  抗血清制备
DOI:
投稿时间:2014-08-07修订日期:2014-09-26
基金项目:科技部动物及动物产品质量安全检测标准物质研制项目禽病病原微生物抗原抗体检测标准物质研制课题(2013FY113300-4);江苏高校优势学科建设工程和江苏省动物重要疫病与人兽共患病防控协同创新中心资助项目 ,陈倩,胡序明,叶建强,邵红霞,钱琨,秦爱建*
Rapid Purification of Egg Drop Syndrome-76 Virus and Preparation of Its polyclonal antibodies
(Key Lab of Jiangsu Preventive Veterinary Medicine of Yangzhou University,Yangzhou)
Abstract:
To make standard antigen and polyclonal antibodies for detection of egg drop syndrome-76 virus (EDS-76), we developed a rapid method of virus purification with standard strain AV-127. Our results showed that the pured virus particles could be obtained by chloroform extraction and PEG concentration without sucrose ultracentrifugation. The HA titer of the purified virus reached to 19Log2. The polyclonal antibodies to EDS-76 generated here showed 13Log2 of HI titer, 1:1000 of IFA titer. Western blot analysis results showed the same pattern to the available standard sera against EDS-76. Moreover, the generated polyclonal antibodies against EDS-76 did not show cross-reaction with other 12 avian pathogens. Overall, our rapid viral purification method provides efficient tools and materials in developing standard antigen and antibodies for detection of EDS-76 in future.
Key words:  Egg Drop Syndrome-76  purification  antiserum generation

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