基因芯片在百合无症病毒检测中的应用

设计百合无症病毒、植物18SrRNA基因的特异扩增引物和各种对照的探针，将探针固定在醛基化玻璃片基上，完成植物基因芯片制备。提取百合种球总RNA，经RT-PCR扩增相应区段并用Cy3dCTP进行标记，用标记的PCR产物与芯片杂交，扫描仪对杂交结果进行扫描，GenePixProd．0软件对杂交图像进行分析。结果从荷兰进口的百合种球中检测出百合无症病毒，证明了该实验研制的植物病毒基因芯片的准确性和灵敏性。     

应用基因芯片检测烟草脆裂病毒

 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.     

从荷兰进口植物中检出南芥菜花叶病毒

南芥菜花叶病毒Arabis mosaic virus(ArMV)是豇豆花叶病毒科Comoviridae线虫传多面体病毒属Nepovirus成员,主要分布在荷兰、比利时等欧洲国家和美国、加拿大、澳大利亚、新西兰、南非、日本,我国尚未有报道。该病毒通常采用血清学和鉴别寄主方法进行检测,但该方法较易出现假阳性,存在检测时间长,准确性和灵敏度较低等问题。近几年发展起来的基因芯片技术已经广泛用于人类病毒如乙肝病毒、人类免疫缺陷病毒、严重急性呼吸系统综合征病毒[1～3]等医学诊断和动物病毒如口蹄疫、禽流感[4]的检测和检疫,而应用基因芯片检测植物病毒实例的报道…     

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。