【摘要】 利用鸡胚成纤维细胞 (CEF)培养禽脑脊髓炎病毒 (AEV) ,经过六次盲传发现 :AEV在 CEF上无细胞病变 (CPE) ,但利用 CEF细胞上清接种 SPF鸡胚 ,可产生不同程度的 AE鸡胚病变。分别取不同时间的感染细胞上清 ,测定 AEV浓度 ,结合培养条件 ,进而确定 AEV培养的最佳时机。结果表明 :以 AEV在 CEF上培养 7天最好 ,病毒滴度可达 10 2 .8EID50 / 0 .2 ml。将经 CEF培养的 AEV差速离心 (浓缩约 5 0 0倍 ) ,接种 SPF鸡胚 ,可产生典型的鸡胚病变 ,其滴度为 8× 10 5.0 EID50 / 0 .2 m l。通过 Cs Cl密度梯度离心提纯病毒 ,在电镜下观察到了大小基本一致的病毒粒子 ,病毒直径约为 2 5 nm。利用 AEV感染的 CEF或通过“细胞飞片”制备荧光片 ,建立了间接免疫荧光快速检验 CEF是否感染 AEV的方法。更多还原

【Abstract】 Avian encephalomyelitis virus (AEV) was blindly passaged in chicken embryo fibroblast (CEF) cell for six generations.We found that: the CEF cell had not CPE expressed,but the SPF chicken embroys inoculated with this CEF cell’s supernatant expressed AE pathological changes. The CEF’s supernatant in different times was respectively collected and detected in order to select the best chance of AEV. Experiment results indicated that: AEV reached the highest titer 10 2.8 EID 50 /0.2 ml after cultivation for 7 days in CEF. The SPF chicken embroys expressed typical pathological changes after cultivation with the condensed AEV for 500 times (AEV density:8×10 5.0 EID 50 /0.2 ml) by different centrifugation.The diameter of the virus was about 25 nm after purification by CsCl density gradient centrifugation. The CEF cultivation infected with AEV was made into fluorescenec slides and indirect immunofluorescent technique (IFA) was set up to detect AEV in the CEF cell culture.更多还原