几种主要禽疫病诊断基因芯片的制备及初步应用  

Preparation and Preliminary Application of Diagnostic Gene Chip of Several Avian Infectious Diseases

作　　者：曹三杰[1] 文心田[1] 肖驰[1] 张焕容[1] 杨利[1] 肖国生[1] 黄小波[1] 

机构地区：[1]四川农业大学动物科技学院动物传染病与基因芯片实验室,动物疫病与人类健康四川省重点实验室,雅安625014

出　　处：《畜牧兽医学报》2006年第4期356-360,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省重点科技攻关项目(01NG018-01);四川省重点学科建设项目(SZD0418)

摘　　要：进行了几种主要禽疫病诊断基因芯片制备及其初步应用研究。试验分别设计和克隆鉴定了NDV、IBV、AIV和IBDV的靶基因重组质粒。以克隆的靶基因重组质粒为模板,分别进行PCR扩增制备靶基因并纯化,以基因芯片点样仪将制备的靶基因点制在氨基化的基片上,经干燥、水合、紫外线交联和洗涤后,成功制备了NDV-IBV-AIV-IBDV诊断基因芯片。试验应用CY3荧光标记制备的探针进行芯片的检验,结果表明制备的NDV-IBV-AIV-IBDV诊断基因芯片质量好,可对NDVI、BV、AIV和IBDV进行诊断检测,具有检测灵敏性好,特异性高和芯片可重复检测的优点。试验对30个临床样品进行初步应用检测,结果表明该诊断基因芯片技术与RT-PCR检测技术检出率基本一致,并具有同步诊断检测多种疫病的优点。Preparation and preliminary application of diagnostic gene chip of several avian diseases have been developed in this paper. Recombinant plasmids of target genes has been designed and cloned respectively. The target genes of NDV, IBV, AIV and IBDV were amplified by PCR with recombinant plasmids templates respectively and purified. Target genes were spotted on glass slides with microarray printing system, and the spotted slide was dried at room temperature, hydrated, lighted with UV and washed. The diagnostic gene chip for detecting NDV, IBV, AIV and IBDV were produced successfully. The results of qualification test showed that the diagnostic gene chip are reliable and well for detecting NDV, IBV, AIV and IBDV, and the sensitivity, specificity and repeatability of this new technology is well for detecting. 30 clinical samples were detected by gene chip technology and RT-PCR at the same time, the results illustrated that the detecting rate for NDV, IBV and IBDV are nearly same between the two ways, but the gene chip technology can detect different disease simultaneously.

关 键 词：鸡新城疫 鸡传染性支气管炎 禽流感 鸡传染性法氏囊病 基因芯片 诊断 

分 类 号：S854.43[农业科学—临床兽医学]