传染性法氏囊病病毒在鸡胚成纤维细胞中的增殖规律  

Proliferation of Infectious Bursal Disease Virus in Chicken Embryo Fibroblasts

作　　者：王小楠[1] 张焕容[1,2] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]动物医学四川省高等学校重点实验室,四川成都610041

出　　处：《动物医学进展》2014年第6期71-76,共6页Progress In Veterinary Medicine

基　　金：西南民族大学研究生创新性科研项目(CX2013SZ73)

摘　　要：为建立传染性法氏囊病病毒(IBDV)弱毒鸡胚成纤维细胞(CEF)感染模型,用10日龄鸡胚制备原代CEF,经过继代培养制备继1代CEF,接种IBDV弱毒后观察细胞病变情况,采用SYBR GreenⅠ实时荧光定量反转录PCR(qRT-PCR)方法检测IBDV在继1代CEF中病毒拷贝数并绘制病毒增殖曲线。结果显示,IBDV接种继1代CEF后,36h开始出现细胞病变,60h时出现明显的细胞病变,表现为细胞间隙增宽,细胞变圆聚集和细胞大量脱落。荧光定量RT-PCR检测感染CEF 12、24、36、48、60、72、96、120、156h的病毒培养液,结果显示病毒拷贝数分别为1.70×107、3.72×107、7.41×107、1.20×108、4.80×107、1.51×107、9.33×106、7.76×106和2.24×106 copies/μL,其中CEF感染IBDV 48h时病毒拷贝数最高。结果表明,成功建立了IBDV弱毒CEF感染模型,为进一步研究IBDV的致病机理、病毒与细胞的相互作用以及疫苗研究奠定了基础。The aim of the study was to establish an infectious bursal disease virus （IBDV ）infection model in chicken embryo fibroblasts （CEF）.CEF were prepared with 10-day old chicken embryo and passaged once.The cytopathic effect （CPE）of IBDV in once-passaged CEF was observed and the dynamic growth of IBDV in CEF was detected by real-time reverse-transcriptase PCR （RRT-PCR）based on SYBR Green Ⅰ. The results showed that CPE first appeared at 36 h post IBDV infection,and the most obvious CPE such as the cell gap increasing,net-balloon filamentation and exfoliated CEF appeared at 60 h post IBDV infec-tion.The real-time RT-PCR analysis showed that the viral RNA loads of IBDV were 1.70×10^7 ,3.72× 10^7 ,7.41×10^7 ,1.20×10^8 ,4.80×10^7 ,1.51×10^7 ,9.33×10^6 ,7.76×10^6 and 2.24×10^6 copies per mi-croliter in cultured virus suspension collected at 12,24,36,48,60,72,96,120 and 156 h post IBDV in-fection,respectively,the highest amount appeared at 48 h.The results indicated that we successfully es-tablished the IBDV infection model in CEF which is useful for further studies concerning IBDV pathogene-sis,virus-cell interaction and vaccine research.

关 键 词：传染性法氏囊病病毒 鸡胚成纤维细胞 病毒增殖 

分 类 号：S852.659.4[农业科学—基础兽医学]