葡萄斑点病毒的血清学检测技术  

Serological testing technique of the grapevine fleck virus

作　　者：刘永清[1] 洪霓[2] 王国平[2] 

机构地区：[1]湖北民族学院生命科学院,湖北恩施445000 [2]华中农业大学植物科技学院,武汉430070

出　　处：《果树学报》2005年第3期289-291,共3页Journal of Fruit Science

摘　　要：应用A蛋白酶联免疫吸附法(ProteinASandwichEnzymeLinkImmunoSorbantAssay,PAS-ELISA)、三抗体夹心酶联免疫吸附法(TripleAntibodySandwich,TAS-ELISA)、酶联板直接捕获抗原的酶联免疫吸附法(PlateTrappedAntigen-Indirect-ELISA,PTA-I-ELISA)、直接组织印迹法(DirectTissueBlotImmunoassay,DTBIA)和圆点免疫印迹分析法(Dot-ImmunoBindingAssay,DIBA)等5种方法检测葡萄斑点病毒(GrapevineFleckVirus,GFkV)感染的葡萄枝条、叶柄和叶片。结果表明:TAS-ELISA法灵敏度最高,其粗汁液的最大稀释度为1:5120,DTBIA和DIBA法均为1:1280,PAS-ELISA法为1:640,PTA-I-ELISA法检测的灵敏度最低,为1:320。针对不同季节和葡萄不同的取样部位进行的血清学检测结果表明,以秋末冬初和葡萄中下部枝条为GFkV最佳的检测时机和器官。另外,用健康汁液吸附抗血清,可以明显降低DTBIA和DIBA的非特异性反应,便于得到直观、准确可靠的检测结果。In this article the GFkV infected grape twigs,petioles and blades were detected by the following 5 methods: Protein A Sandwich Enzyme Link ImmunoSorbant Assay (PAS-ELISA),Triple Antibody Sandwich- ELISA (TAS-ELISA), Plate Trapped Antigen-Indirect-ELISA(PTA-I-ELISA) , Direct Tissue Blot Immunoassay (DTBIA) and Dot-ImmunoBinding Assay (DIBA).The findings showed that the TAS-ELISA method was the most sensitive one, its maximum dilute folds of the extract was 1=5120, and both DTBIA and DIBA method were 1 = 1280, and PAS-ELISA method was 1:640, while the PTA-I-ELISA detecting had the minimum sensitivity, only 1:320. The results of serological detection of different parts of the grapevine in different seasons showed that the middle-lower twigs of grape vines were the best materials for detecting GFkV, and the optical time was at the end of autumn and in early winter. Furthermore, using the healthy extract to absorb serum could obviously decrease the non-specific reaction of DTBIA and DIBA, so that the clear, accurate and reliable result can be achieved.

关 键 词：葡萄斑点病毒 血清学检测 

分 类 号：S663.1[农业科学—果树学]