10种植物病毒的基因芯片检测技术研究

 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。

Technique for detection of 10 plant viruses by gene chip

The probes of 10 plant viruses and control were designed and immobilized on chips.The total RNA was extracted by Trizol method from plants.Primers were designed according to the sequences of cp gene or rep gene of the plant viruses.The corresponding fragments were amplified by RT-PCR and the PCR products were labeled by Cy3-dCTP.The labeled PCR products were hybridized with the probes and the signals were scanned by scanner and analyzed by GenePix Pro 4.0 software.Specific signals were identified from virus-infected plants.The detection sensitivities of gene chip were 10-100 times higher than RT-PCR.The results indicate that gene chip can be used for detection of plant viruses fast and accurately.