禽流感H5和H9亚型基因探针的制备  

Preparation of the Probes Used for H5 and H9 Subtype of Avian Influenza Virus

作　　者：方浩[1] 宋念华[2] 姚志伟[1] 熊元林[1] 王松太[1] 

机构地区：[1]武汉生物工程学院生物工程系,武汉430415 [2]湖北省动物疫病诊断中心,武汉430064

出　　处：《湖北农业科学》2007年第2期190-193,共4页Hubei Agricultural Sciences

基　　金：湖北省科技攻关项目(2004AA202B04)

摘　　要：参照GeneBank数据库上H5N1和H9N2 AIV的HA基因序列,选取保守区域,设计了两对引物,分别用于扩增HA基因,产物纯化回收得到180bp和285bp cDNA片段,采用随机引物法Digxigenin-11-dUTP标记得到探针H5PR和H9PR,通过斑点杂交检测,结果表明探针的灵敏度高和特异性好,可以进行相关性试验。Based on conserved sequences of HA gene of H5N1 and H9N2 selected in Genebank database, two pairs of primers were designed and used to amplify the conserved sequences. Two cDNA fi＇agments of about 180bp and 285bp size were obtained after purifying the products. Then the probes named H5PR and H9PR were obtained by labeled individually the two fragments with Digxigenin-11-dUTP using the random primed method. Through detecting the sensitivity and specificity of the probes with dot blot experiments. The results showed the probes had good performance, and could be used for some correlative experiments.

关 键 词：禽流感病毒 斑点杂交 地高辛探针 制备 H5亚型基因 H9亚型基因 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]