AIV血凝素亚型同步检测基因芯片技术研究  

Simultaneity Detection of HA Subtypes of Avian Influenza Virus on DNA Microarray

作　　者：王秀荣[1] 刘丽玲[1] 包红梅[1] 乔传玲[1] 王煜[1] 孙凯[1] 孔宪刚[1] 陈化兰[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《生物技术通报》2006年第C00期260-263,271,共5页Biotechnology Bulletin

基　　金：国家十五重点攻关项目:不同动物流感病毒免疫学及高通量检测技术的研究和开发(2004BA519A23);国家自然科学基金项目"应用cDNA芯片技术研究禽流感病毒毒力相关基因表达差异"(30200201;3044009)

摘　　要：对流感病毒14个血凝素亚型的基因芯片检测技术进行了初步研究。通过RT-PCR克隆禽流感病毒血凝素基因片段,获得重组质粒。从重组质粒扩增大约500bp的DNA片段,浓缩后点到氨基化玻璃载体上,制成芯片。待检病毒样品用TRIzolLS提取RNA,反转录过程中用Cy5标记样品cDNAs。将标记样品与芯片杂交,扫描芯片上待检样品与芯片上捕捉探针的结合位点,杂交信号与预期设想一致。结果显示,DNA芯片技术可以提供一种有效的AIV血凝素亚型鉴别诊断方法。In this study, a rapid microarray-based assay for the reliable detection of HA subtypes of avian influenza virus （AIV） was preliminary developed. The cDNA clones which encode approximately 500bp avian influenza virus gene fragments were obtained by reverse-transcription （RT） PCR technique, the DNA fragments which were reamplified from recombinant plasmids, were spotted on a glass-bound microarray, Cy5-1abeled fluorescent probes were hybridized to these sample cDNAs, which generated from viral RNA during RT. The arrays were scanned to determine the probes binding sites. The hybridization pattern agreed approximately with the known grid location of each target. The results showed that DNA microarray technology can provide an useful diagnostic method for AIV.

关 键 词：禽流感病毒 基因芯片 血凝素亚型鉴定 AIV 

分 类 号：Q789[生物学—分子生物学]