新型基因芯片检测禽流感病毒、鸡新城疫病毒和鸡减蛋综合征病毒  

Detection of AIV,NDV and EDSV with a New Type Gene Chip

作　　者：廖秀云[1] 陶虹[2] 邵建宏[1] 罗宝正[1] 薄清如[1] 刘国昌[3] 徐海聂[1] 沙才华[1] 

机构地区：[1]珠海出入境检验检疫局,广东珠海519015 [2]深圳出入境检验检疫局,广东深圳518045 [3]华南农业大学兽医学院,广东广州510642

出　　处：《动物医学进展》2013年第10期15-20,共6页Progress In Veterinary Medicine

基　　金：国家质检总局科技计划项目(2009IK013)

摘　　要：建立了可同时筛选禽流感病毒(AIV)通用型、H4、H5、H6、H7、H9、N1、N2、N9亚型及鸡新城疫病毒(NDV)、鸡减蛋综合征病毒(EDSV)的基因芯片检测方法。从GenBank中下载相关序列,利用分子生物学软件设计特异性引物和杂交探针。使用Dr.chip系统进行芯片点样、杂交、显色、读数,优化反应条件后做灵敏度、特异性试验,然后应用于临床检验。结果显示,各病原间无交叉反应,探针可特异性识别靶基因;病原的最低浓度为4.83pg/μL,灵敏度较高;每个梯度的两个反应结果一致,重复性良好。对临床样品检验,结果同病毒分离鉴定方法一致。本研究建立的基因芯片检测方法可用于禽流感通用型、H4、H5、H6、H7、H9、N1、N2、N9亚型及NDV、EDSV的鉴别检测。This study aimed to establish a new type gene chip detection method that can screen the avian in- fluenza virus （AIV） generic type and identificate H4, H5, H6, H7, H9, N1, N2, N9 subtypes, Newcastle disease virus （NDV） and egg drop syndrome virus （EDSV） simultaneously. Specific primers and hybridization probes were designed by molecular biology software after the relevant sequences were obtained from GenBank. Dr. chip system was used for chip spotting, hybridization, coloring and reading data, opti- mized the reaction conditions before sensitivity experiment and specificity experiment, then applied to the clinical examination. Hybridization results displayed that no cross-reactivity between different pathoge- nies, probes can specifically recognize the target gene~ The minimum concentration to detect these patho- gens is 4.83 pg/tzL and has high sensitivityl Two reaction results in each gradient are consistent and have favorable reproducibility. The results of detecting clinical samples were consistent with the virus isolation method. The gene chip detection method established in this study can be used for differential detection of influenza virus （AIV） generic type and identificate H4, H5, H6, H7, H9, N1, N2, N9 subtypes, NDV and EDSV.

关 键 词：基因芯片 禽流感病毒 新城疫病毒 鸡减蛋综合征病毒 

分 类 号：S852.659.5[农业科学—基础兽医学]