应用两种基因组快速扩增方法进行病毒芯片杂交鉴定  

Evaluation of Two Rapid Genome Amplification Methods to Identify Virus Genus by Microarray Detection

作　　者：康晓平[1] 刘洪[1] 杨银辉[1] 胡玉洋[1] 朱晓光[1] 祝庆余[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国生物化学与分子生物学报》2006年第9期750-754,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家科技攻关计划项目资助(No.2003BA712A01)~~

摘　　要：为了摸索均衡的病毒基因组扩增方法,建立高通量的病毒检测基因芯片技术平台,本研究以甲病毒属的辛德比斯病毒作为检测模型,分别以随机PCR扩增法和MDA(multipledisplacementamplification)扩增法扩增病毒基因组,并以两种扩增产物作为模板,扩增辛德比斯病毒的特异基因片段以初步验证基因组扩增的均衡性;然后将两种基因组扩增产物标记荧光染料后与基因芯片进行杂交;结果表明从两种基因组扩增产物中正确扩增出了辛德比斯的特定基因片段,作为探针可与基因芯片上的靶标基因特异性结合;基因组扩增产物与基因芯片进行杂交,可成功检测到甲病毒属的特异性信号,充分说明随机PCR扩增法和MDA扩增法扩增病毒基因组可用于病毒性病原体的基因芯片检测.To develop proper method for unbiased amplification of viral genome, and exploit strategy for highly parallel viral screening by microarray-based detection. The genome of sindbis virus was amplified by random PCR and MDA strategies respectively, and the products were used as template to amplify specific gene in it to test the quality of the two genome amplification method ; then the genome amplification products of sindbis virus were labeled with fluorescence dye and detected with microarray. Results indicated that the specific gene in the genome of sindbis virus was correctly amplified out from the products of the two genome amplification method, and specially bound to the target gene by microarry detection. Applying the genome amplification products of sindbis virus to microarry detection, signals of alphavirus were detected successfully, which suggested that both random PCR and MDA could amplify the genome of virus and be applied to microarray-based detection of viral pathogens.

关 键 词：病毒 基因组扩增 芯片检测 

分 类 号：R394.2[医药卫生—医学遗传学]