Total RNA in tulips was extracted by Trizol method.Primers were designed according to the sequences of Tobacco rattle virus and 18S rRNA gene of plant.The corresponding sections were amplified by RT-PCR and the PCR products were labeled by Cy3-dCTP.The probes of plant virus,18S rRNA gene and comparisons were designed and immobilized on chips.Labeled PCR products were hybridized with the probes and the signals were scanned by scanner and analyzed by GenePix Pro 4.0 software.Tobacco rattle virus was detected from tulips which were imported from Holand.The accuracy and sensitivity of the plant virus gene chip were proved.
本研究设计了10种植物病毒和各种对照的探针，将探针固定在醛基化玻璃片基上制备基因芯片。用常规的Trizol法提取植物总RNA，根据病毒的外壳蛋白基因或复制酶基因设计特异性引物，经RT—PCR扩增相应区段并用Cy3-dCTP进行标记。将标记的PCR产物与芯片杂交，扫描仪对杂交结果扫描，GenePix Pro 4．0软件对杂交图像进行分析。结果表明，该芯片可以从病毒感染样本中检测到特异性识别信号，检测灵敏度比RT—PCR高10-100倍，所以，基因芯片能对植物病毒作出快速、准确的检测。
