痘苗病毒寡核苷酸检测芯片的设计及研制  

Design and preparation of oligonucleotide microarray for vaccinia virus detection

作　　者：王艳[1] 马文丽[1] 毛向明[1] 吴清华[1] 李凌[1] 王洪敏[1] 肖维威[1] 郑文岭[2] 

机构地区：[1]第一军医大学分子生物研究所,广东广州510515 [2]广州军区广州总医院医学实验科,广东广州510010

出　　处：《第一军医大学学报》2004年第2期180-183,共4页Journal of First Military Medical University

基　　金：全军十五重大攻关项目~~

摘　　要：目的对痘苗病毒进行寡核苷酸检测芯片的初步研究,为建立寡核苷酸芯片检测该类病毒提供初步研究依据。方法根据痘苗病毒特异基因设计寡核苷酸探针,人工合成探针后制备寡核苷酸芯片。在病毒感染的不同阶段提取病毒样品DNA及阴性样品DNA,采用限制性显示技术标记,标记样品与芯片杂交后,用Agilent芯片扫描仪检测杂交结果。结果芯片与病毒样品杂交有较强的杂交信号,而与阴性样品杂交除阳性探针外均无信号。结论病毒样品与阴性样品杂交信号区别明显,在病毒感染的各个时段也都有明显的杂交信号,反映了寡核苷酸芯片具有较高的特异性和灵敏度。Objective To study the preparation of oligonucleotide microarray for detecting vaccinia virus. Methods Oligonucleotide probes were designed and synthesized according to the specific genes of vaccinia virus. Sample DNA of the virus and the negative control sample were obtained and labeled by restriction display technique, followed by hybridization to the oligonucleotide microarray and scanned by Agilent scanner. Results Strong hybridization signals were detected from the viral DNA hybridized with the microarray, but were absent in the negative sample when positive probes were not used. Conclusion Distinct differences in the hybridization signals between the virus sample and negative sample and between the samples obtained in different phase of infection demonstrate high specificity and sensitivity of the microarray for vaccinia virus detection.

关 键 词：寡核苷酸 设计 痘苗病毒 核酸探针 DNA芯片 检测 疫苗 天花 

分 类 号：R346[医药卫生—基础医学]