百合病毒的DNA芯片检测技术研究

Detection for CMV,LSV,LMoV Infected Lily with DNA Microarry Techniques

根据已知的黄瓜花叶病毒,百合无症病毒、百合斑驳病毒基因核苷酸序列,设计引物和探针,制备寡核苷酸芯片。用Cy3标记核苷酸引物,不对称RT-PCR扩增产物与芯片上的寡核苷酸探针杂交,荧光扫描仪检测并分析信号。研究制备的基因芯片能够检测侵染百合的3种重要病毒核酸的特异性荧光信号,该项技术具有特异、灵敏、快速的优点。

Specific primers and probes were designed according to conservative and specific DNA sequence of CMV,LSV and LMoV. The probes were spotted on glass slides to form microarrays The Cy3-labled single strand DNA fragments prepared by dissymmetical RT-PCR were hybridized with the probes on the glass slides . The microarrays were scanned and analyzed with a scanner. DNA microarray could detect different typed DNA of CMV, LSV and LMoV with adequate specificity and sensitivity. Microarray techniques establish a rapid,sensitive and specific diagnostic assay for lily viruses. The successfully developed DNA microarray and techniques might be a very useful method for diagnosis and prevention ,and widely applied in specific pathogen detection of diseases such as lily virus.