摘要
以香蕉交脉蚜（Ｐｅｎｔａｌｏｎｉａｎｉｇｒｏｎｃｒｖｏｓａ）人工接种表现典型束顶病症状的香蕉组培苗为材料，预预先经氯仿－正丁醇充分乳化的缓冲液抽提和澄清、差速离心、蔗糖垫部分纯化、蔗糖梯度离心，得到粒体完整的直径约１８－２０ｎｍ的球状病毒。提纯的病毒制剂具典型的核蛋白紫外吸收光谱，最高吸收峰在２５７ｎｍ左右，最低吸收峰在２４０ｎｍ左右，Ａ２６０／２８０＝１．３，提纯产量最高可达３．４ｍｇ／ｋｇ组织。用上述提纯病毒作为抗原免疫家兔制备的抗血清，经对流免疫电泳方法测定效价为１：３２。用该抗血清建立的Ａ蛋白夹心ＥＬＩＳＡ（ＤＡＳ－ＥＬＩＳＡ）及与单克隆抗体结合使用的异种抗体双夹心ＥＬＩＳＡ（ＤＡＳ－ＥＬＩＳＡ）能检测各种香蕉束顶病样品。
Isometric virus particles，18－20nm in diameter，were purmedfrom micropropagatedbanana plants with typical bunchy top via viruliferous Pentalonia nigronervosa inoculation，byclarifying with chloroformbutanol，concentrating with one cycle of different centrifugation，cen-trifugating on the top of a twolayer sucrose cushion of 10％ and 30％ and then on sucrose densitygradient．The U V absorbance profile of the preparation was charateristic of a nucleoprotein hav-ing a maximum absorbance at 257nm，a minimum absorbance at 240nm and an A260／280 ratioof about 1．3．The highest yield was up to 3．41 mg／kg infected tissue. An antiserum with avirus-specific titre of1／32 by immunoelectrophoresis was obtained by immnunizing a rabbit usingthe purified virus preparation. The polyclonal antiserum was successfully used in a protein Asandwich ELISA（PAS-ELISA）and a double antibody sandwich-EIISA（DAS-ELISA）with amonoclonal antibody to detect BBTV in various disease samples.
出处
《热带作物学报》
CSCD
1996年第2期42-46,共5页
Chinese Journal of Tropical Crops
基金
福建省热带作物研究所自选课题
关键词
香蕉
束顶病毒
提纯
血清学
Banana bunchy top virus（BBTV）；Purification；Serology；Protein A sandwich ELISA（PAS-ELISA）
Double antibody sandwich ELISA（DAS-ELISA）
