葡萄斑点病毒的血清学检测技术Serological testing technique of the grapevine fleck virus
刘永清,洪霓,王国平
摘要(Abstract)：
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。
关键词(KeyWords)： 葡萄斑点病毒;血清学检测
基金项目(Foundation):
作者(Author): 刘永清,洪霓,王国平
Email:
DOI:
参考文献(References)：
- [1] Edwards ML,Cooper JI.Plant virus detection using a new form of indirect EUSA[J]. J Virol Meth, 1985,11: 309-319.
- [2] Cambra M, Asensio M,Gorris MT, et al. Detection of plum pox virus using monoclonal antibodies to structural and non -structural pro-teins[J]. Bulletin OEPP/ EPPO Bulletin,1995,24:569-577.
- [3] Faggioli F, Serrone PD, Barba M. Spread of grapevine fleck virus (CFkV)in latium and its rapid serological detection[J]. Rivista di viticotura e di Enologia,1993,46(2): 55-59.
- [4] Permar TA, Gamsey SM, Hendersom CT. Direct tissue blot immunoas-says for detection of citrus tristeza virus (CTV)[J]. Phytopathology, 1998,82:60.
- [5] Wang Q, Shi Y, Yang J, et al. Detection of apple chlorotic liaf spot virus and apple stem grooving virus by Dot-immunobinding assay[J]. Acta Hort,1998, 472:51-54.
- [6] Walter B, Bass P,Legin R. The use of a gveen-grafting technique for the detection of virus-like disease of the grapevine[J]. PHytopathol, 1990,128:137-145.