口蹄疫等5种动物病毒基因芯片检测技术的研究  

Study on Diagnostic Gene Chip for Detecting Five Animal Viruses

作　　者：杨素[1] 花群义[2] 徐自忠[2] 杨晶焰[2] 谭德勇[3] 董俊[2] 杨云庆[2] 赖建华[3] 

机构地区：[1]珠海出入境检验检疫局,珠海519015 [2]云南出入境检验检疫局,昆明650228 [3]云南大学生命科学学院生物化学与分子生物学实验室,昆明650091

出　　处：《微生物学报》2004年第4期479-482,i001,共5页Acta Microbiologica Sinica

基　　金：国家科技攻关重大专项资助项目 ( 2 0 0 1BA80 4A2 2 ) ;云南省科技攻关计划重点项目 [云计科技 ( 98) 63 6号 ] ;昆明市科技计划项目[昆科 (农 )字 2 0 0 2 0 2 0 0 3 ]~~

摘　　要：用分子克隆方法获得口蹄疫病毒、水泡性口炎病毒、蓝舌病病毒、鹿流行性出血热病毒和赤羽病病毒各一段高度保守的基因片段 ,用芯片点样仪点样到包被过的玻璃片上 ,制备成检测芯片。提取样品中的RNA ,进行反转录和荧光标记后滴加到芯片上进行特异性杂交 ,对杂交结果进行扫描检测 ,可同时诊断上述 5种动物传染病 ,此方法不但快速、准确、敏感 ,而且可同时进行多种病毒的检测 。A gene chip was developed by preparing highly conserved DNA fragments for five animal infectious disease viruses ( Foot and mouth disease virus, Vesicular stomatitis virus, Bluetongue virus, Epizootic haemorrhagic disease virus of deer, Akabane disease virus ) using molecular cloning methods, and the DNA fragments were spotted on to specially treated glass slides. RNA extracted from samples was reverse transcripted, followed by PCR amplification and labeling with fluorescent dyes. The labeled cDNA and prepared gene chip were subjected to specific hybridization, and the hybridization results were detected by photoscanning. The results showed that this gene chip technique could identify and distinguish the five animal infectious diseases viruses. This method is rapid, sensitive and specific, and can screen and quarantine a large number of animal samples within a very short period of time.

关 键 词：口蹄疫 动物病毒 基因芯片 分子克隆 水泡性口炎病毒 蓝舌病病毒 鹿流行性出血热病毒 

分 类 号：S854.4[农业科学—临床兽医学]