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应用液氮冷冻香蕉束顶病组织,高速粉碎,氯仿—正丁醇澄清,差速离心,沉淀悬浮后经蔗糖垫部分纯化病毒,收集的蔗糖垫浓缩后经0—40％连续蔗糖梯度离心4小时,在离心管上半部形成一个明显的含有大量等径病毒的紫外吸收峰。再经不连续硫酸铯梯度离心6小时,形成一狭窄的乳白色带,含有高度纯化的直径约18nm的香蕉束顶病毒(BBTV)。用台湾提供的BBTV单克隆抗体,ELISA法可检测到提纯过程中病毒的存在,ISEM法可从粗提物中捕获大量等径病毒。ELISA法也用于检测田间及组织培养原种苗的带毒情况。
The supernatant tissues of banana bunch were frozen by liquid nitrogen, purified by high-speed pulverization, clarified by chloroform-n-butanol, differentiated by centrifugation and partially precipitated by sucrose pad. The collected sucrose pads were concentrated and then subjected to gradient centrifugation 0-40% sucrose 4 H, an obvious UV absorption peak containing a large amount of isoniae virus was formed in the upper half of the centrifuge tube. Centrifugation through a discontinuous cesium sulfate gradient for 6 hours resulted in a narrow milky band containing highly purified banana bunchy top virus (BBTV) of about 18 nm in diameter. The BBTV monoclonal antibody provided by Taiwan was used to detect the presence of virus in the purification process by ELISA. ISEM method can capture a large amount of iso-virus from the crude extract. ELISA method is also used to detect the field and tissue culture of seedlings with poisoning.