H5N1亚型禽流感病毒和新城疫病毒感染鸡胚成纤维细胞IFN-β基因的动态表达

The Dynamics of IFN-β mRNA Expression in Chick Embryo Fibroblast Infected with H5N1 Avian Influenza Virus and Virulent Newcastle Disease Virus

根据鸡β-干扰素（ChIFN-β）基因保守序列设计引物建立了检测鸡IFN-βmRNA表达水平的荧光定量RT-PCR（RRT-PCR）方法,并对H5N1亚型禽流感病毒（AIV）和新城疫病毒强毒株（vNDV）感染后3、6、12、24、30 h的鸡胚成纤维细胞（CEF）中IFN-βmRNA表达水平进行了检测。结果显示,建立的RRT-PCR特异性好,对鸡IFN-βmRNA的扩增效率为94.18%,线性范围为10-8-10-3,相关系数为0.992,最低能检出48拷贝/反应。AIV在感染CEF后6、12 h显著抑制IFN-βmRNA的表达,24 h开始诱导IFN-βmRNA表达,30 h时IFN-βmRNA水平显著升高;vNDV在感染CEF后的24 h内显著抑制IFN-βmRNA的表达,30 h时则显著诱导IFN-βmRNA表达。本试验结果为进一步研究H5N1和NDV与机体的相互作用提供了有价值的信息。

According to the chicken＇s IFN-β（ChIFN-β） gene sequences,a pair of primers was designed for developing a SYBR Green I quantitative real-time reverse-transcription PCR（RRT-PCR） to detect the IFN-β gene of chicken,and the expression levels of IFN-β mRNA in chick embryo fibroblast（CEF） at 3,6,12,24 and 30 h after infected with H5N1 avian influenza virus（AIV） and virulent Newcastle disease virus（vNDV） were detected.The results showed that the assay was specific for ChIFN-β gene with amplification efficiency of 94.18%,and the linear range was 10-8 to 10-3 with good correlation coefficient of 0.992,as well as the detection limit reached to 48 copies per reaction.The expression of IFN-β mRNA in CEF infected with AIV H5N1 and vNDV showed,the expression of IFN-β mRNA in CEF was suppressed significantly at 6 and 12 h,then induced at 24 h and significantly induced at the 30 h after infected with AIV H5N1;the IFN-β mRNA expression of CEF was suppressed significantly at 24 h and induced significantly at 30 h after infected with vNDV.The results provided valuable information for further researching the interaction of H5N1/NDV and organism.