以葡萄扇叶病毒(GFV)干u马铃薯Y病毒(PVY)外壳蛋白基因的重组质粒为模板,用聚合酶链式反膨技术(PCR)分别合成了长度为1.5kb和0.75kb的生物素标记双链cDNA探针,在斑点杂交反应中,探针的最适使用浓度为1/100,GFV探针检测GFV-RNA的灵敏度为10pg,检测提纯GFV的灵敏度为25pg,检测感病苋色藜的最大稀释倍数为10000倍;PVY探针检测PVY-RNA的灵敏度为30pg,检测提纯PVY灵敏度为500pg,感病烟草检测的最大稀释度为8000倍,阴性对照均无杂交信号出现。

Both grapevine fanieaf virus and potato virus Y biotin-labelled ds-cDNA probes were synthesized by polymerase chain reaction using recombinant coat protein gene as template. The optimum concentration of ds-cDNA probes were 1:100 in dot blot hybridization. The sensitivity of detection for GFV-RNA was 10 pg,partially purified GFV was 25pg,the maximum dilution of GFV-infected Chenopodium amaranticolor leaves extract was 1:10000; The sensitivity of detection for PVY-RNA was 30pg, partially purified PVY was 500pg,the maximum dilution of PVY infected Nicotiana tabacum leaves extract was 1:8000. There were no any hybridization signales in the negative control.