鸡传染性支气管炎病毒M41型的扩繁及在鸡胚肾细胞上的适应性研究  

On propagation and adaptability of avian infectious bronchitis virus-M41 in chicken embryo kidney cells

作　　者：李华伟[1] 武聚富[1] 庄英华[1] 高飞[1] 吴国娟[1] 

机构地区：[1]北京农学院动物科学技术学院,北京102206

出　　处：《北京农学院学报》2011年第3期31-34,共4页Journal of Beijing University of Agriculture

基　　金：国家自然科学基金项目(30771625;30972215);北京市自然科学基金项目(6092004);北京市属市管高校人才强教计划项目(PHR2009-07135)

摘　　要：探明鸡传染性支气管炎病毒(Avian Infectious Bronchitis Virus,IBV)感染鸡胚肾细胞(Chicken Embry-o Kidney Cells,CEK cell),并且实现其增殖为目的。将IBV稀释液注射到SPF鸡胚尿囊腔内,培养鸡胚扩繁,观察记录结果;并培养CEK细胞,采用100倍半数组织培养感染量的IBV进行染毒实验;应用Reed-Muench法计算半数鸡胚感染量为10-6.68/0.1 mL;用Real time PCR检测IBV在感染细胞中的含量,以确定IBV在CEKcells中已经感染且增殖成功。结果表明,接种10日龄的SPF鸡胚可以收集更多的、约8 mL的尿囊液毒;反复盲传至7代的IBV可使CEK cells产生明显的细胞病变效应;通过实时荧光定量PCR和RT-PCR检测,最终确定IBV-M41型扩繁成功并且能够在CEK细胞上繁殖,为下一步实验提供了参考。This study investigated the avian infectious bronchitis virus（IBV） –M41 infected chicken embryo kidney cells（CEK cell） and could multiply successfully.The IBV was injected into the SPF chicken eggs to propagate.And CEK cells were cultured ahead of adding in the IBV of 100TCID50.Reed-Muench was used to calculate the EID50.The real time PCR assay was used to detect the content of IBV in CEK cells whitch past infection.IBV-M41 was determined and it multiplied in CEK cells successfully.The results showed that the 10-day-old SPF chicken embryos inoculated can collect more of allantoic fluid virus（about 8 mL）;the CEK cells infected with 7th generation of IBV can produce significant cytopathic effect;and ultimately determined by quantitative real-time PCR and RT-PCR assay,the propagation of IBV-M41 and the ability to reproduce in CEK cells was succesfful.The results of this study provides a reference for further experiments.

关 键 词：传染性支气管炎病毒 鸡胚肾细胞 PCR 适应性 

分 类 号：S852.6[农业科学—基础兽医学]