AIV血凝素亚型同步检测基因芯片技术的研究.pdfVIP

生物技术通报 2006 BIOTECHN OLOG Y B ULLETIN A IV 王秀荣 刘丽玲 包红梅 乔传玲 王煜 孙凯 孔宪刚 陈化兰 ( , 150001) : 对流感病毒 14 个血凝素亚型 基因芯片检测技术进行了初步研究通过 RTPCR 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 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 TR Izol LS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 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 A IV 血凝素亚型鉴别诊断方法 : 禽流感病毒 基因芯片 血凝素亚型鉴定 Simultaneity Detection of HA Subtypes of A vian Influenza V irus on DNA M icroarray W a g X iuro g Liu L ili g Bao Ho gmei Q iao Chua li g W a g Yu Su K ai Ko g X ia ga g Che Huala (A nmi al Inf luenza Laboratory of M inistry of Agriculture and Na tionalK ey Laboratory of Veterinary B iotechnology of H arbin Veterinary R esearch Institu te of CAA , Harbin 150001) Abstract: I this study, a rap id m icroarraybased assay for the reliab le detectio of HA subtypes of avia i flue za virus (AIV ) was prelmi i ary developed. The cDNA clo esw hich e code approxmi ately 500bp avia i flue za virus ge e fragm e tsw ere obtai ed by reversetra scriptio ( RT) PCR tech ique, theDNA fragm e tsw hich were reamp lified from re comb i a t p lasm ids, w ere spotted o a glassbou d m icroarray. Cy5labeled fluoresce t probes w ere hybrid ized to these sam ple cDNAs, wh ich ge erated from viral RNA duri g RT. The arrays were sca ed to determ i e the probes bi di g sites. The hybridizatio patter agreed approxmi ately w ith the k ow grid locatio of each target. The results showed that DNA m icroarray tech ology ca provide a useful diag ostic m ethod for AIV. Key w ords: Avia i flue za virus DNA m