Tannock G A, Shafren D R
Avian Dis. 1985 Apr-Jun;29(2):312-21.
A rapid procedure for the purification of egg-grown or field preparations of avian encephalomyelitis virus (AEV) of neural origin is described. Extracts of infected tissues were clarified and then partly purified with trichlorotrifluorethane (Freon TF), and the virus present was concentrated with polyethylene glycol. The concentrates were then re-extracted with Freon, and a portion was labeled with 125iodine. During subsequent purification steps, virus could be readily detected by monitoring for radioactivity, thus eliminating the need to determine the infectivity in individual fractions or to examine for the presence of virions by electron microscopy. Final purification was achieved by cesium-chloride equilibrium or sucrose-velocity-gradient centrifugation. Virus purified in this manner was shown to be free of tissue debris, to be specific for AEV by immune electron microscopy, and to possess structural proteins characteristic of picornaviruses.
本文描述了一种快速纯化源自神经组织的禽脑脊髓炎病毒(AEV)的鸡胚培养物或野外分离物的方法。感染组织的提取物经过澄清处理,然后用三氯三氟乙烷(氟利昂TF)进行部分纯化,并用聚乙二醇浓缩其中存在的病毒。然后用氟利昂对浓缩物进行再提取,并将一部分用125碘进行标记。在随后的纯化步骤中,通过监测放射性可轻松检测到病毒,从而无需确定各个组分的感染性或通过电子显微镜检查病毒粒子的存在。最终纯化通过氯化铯平衡离心或蔗糖密度梯度离心实现。以这种方式纯化的病毒显示无组织碎片,通过免疫电子显微镜鉴定对AEV具有特异性,并具有微小核糖核酸病毒的特征性结构蛋白。