为明确沙地葡萄茎痘相关病毒(GRSPaV)在山东省发生的情况,2006-2007年在葡萄主栽区和葡萄品种资源圃进行了田间调查、样品采集和病原检测。应用GRSPaV外壳蛋白基因表达获得的多克隆抗体,经间接酶联免疫吸附试验(PTA-ELISA)和斑点酶联免疫吸附试验(Dot-ELISA)检测,在2a采集的298个样品中,125个样品为阳性反应,阳性率41.95%。125个阳性样品涉及43个品种,品种被侵染率74.14%。研究对检测的样品用RT-PCR法验证,有3对引物均扩增出了预期片段,分别为解旋酶基因340bp片段、外壳蛋白(CP)基因842bp片段和RNA聚合酶(RdRp)基因499bp片段。 In order to clarify the occurrence of GRSPaV in Shandi in Shandong Province, field surveys, sample collection and pathogen detection were conducted in grapevine planting areas and grape variety resource farms in 2006-2007. Polyclonal antibodies against GRSPaV coat protein gene were detected by indirect enzyme-linked immunosorbent assay (PTA-ELISA) and dot-ELISA (Dot-ELISA). Among 298 samples collected in 2a, 125 samples As a positive reaction, the positive rate of 41.95%. Of the 125 positive samples, 43 varieties were involved, and the infected rate of varieties was 74.14%. Three pairs of primers were used to amplify the expected fragments of the tested samples, namely the 340 bp fragment of the helicase gene, 842 bp of the coat protein (CP) gene and 499 bp of the RNA polymerase (RdRp) gene. Fragment.