辣椒环斑病毒提纯及抗血清制备

摘要

为获得高效、灵敏的辣椒环斑病毒(Chilli ringspot virus,ChiRSV)多克隆抗体,通过差速离心、密度梯度离心等方法首次对辣椒环斑病毒进行了分离纯化,并将获得的高纯度病毒粒子用于免疫新西兰大白兔,制备了该病毒的多克隆抗血清.获得的抗血清经间接法酶联免疫吸附测定(ID-ELISA),效价高达1:125 000,且特异性高,与黄灯笼辣椒中另一种亲缘关系很近的同属病毒辣椒脉斑驳病毒(Chilli veinal mottle virus,ChiVMV)不发生血清交叉反应、可有效鉴别这2种Potyvirus病毒.获得的抗血清可用于ChiRSV的检测及后续实验,最适工作浓度在1∶5000～1∶25000之间.%In order to acquire polyclonal antibody of Chilli ringspot virus (ChiRSV), the virus was purified using the methods of differential centrifugation and density gradient centrifugation. The purified virion was injected to a New Zealand rabbit to produce polyclonal antibody. The result of ID-ELISA showed that the titer of the antiserum could reach as much as 1 : 125 000. The antiserum also showed high specificity. It showed no cross reaction with Chilli veinal mottle virus (ChiVMV), which could also infect chilli and belonged to the same genus as ChiRSV. The antiserum could efficiently distinguish these two potyviruses. It could be well applied to detect ChiRSV and further experiments, and the most available dilution was around 1:5 000～1:25 000.