甘蔗花叶病毒的提纯及抗血清制备

Purification of Sugarcane Mosaic Virus and Preparation of an Antiserum against the virus

以ＳｃＭＶ－Ａ为材料，采用ＰＥＧ沉淀结合差速离心技术，经１０－４０％蔗糖密度梯度离心后再经浓缩后即为提纯病毒制剂。该提纯病毒具有典型的核蛋白吸收曲线，最大紫外吸收值在２５７ｎｍ处，最小紫外吸收值为２４０ｎｍ，Ａ２６０／２８０＝１．６８，病毒产量可达１．２５－１．３７ｍｇ／ｋｇ。将提纯病毒免疫家兔制备抗血清，所制备的抗血清经Ａ蛋白酶联免疫吸附法测定，其效价为１／１０２４，且可用于检测甘蔗花叶病毒。

Sugarcane mosaic virus was purified from the leaves of the diseased sweet maize inoculated artificially with ScMV-A and used in the experiment. The procedures of purification involved precipitation with PEG differential centrifugation and 10-40% sucrose density gradient centrifugation. The virus preparation was examined with a UV spectrophotometer and showed a typical nucleoprotein spectrum with maximum absorption at 257 nm and minimum at 240 nm, A260/280=1.68. The yield of ScMV ranged from 1.25 to 1.37mg per kilogramme of diseased leaves. The purified ScMV preparation was injected into rabbits to produce an antiserum to ScMV. Titer of the ScMV antiserum was 1/1024 by SPA-ELISA. The ScMV antiserum obtained can also be used to detect ScMV.