两种出血热病毒新型液相芯片检测体系的建立  

Novel liquidchip array of detection for two kinds of hemorrhagic fever virus

作　　者：杨洪[1,2] 王静[1] 刘衡川[2] 杨宇[1] 杨永莉[1] 张乐[1] 

机构地区：[1]中国检验检疫科学研究院卫检所,北京100123 [2]四川大学公共卫生学院

出　　处：《中国国境卫生检疫杂志》2011年第5期311-314,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：财政部质检公益项目(10-44)

摘　　要：目的建立两种出血热生物恐怖病毒(马尔堡病毒、埃博拉病毒)的新型液相芯片检测方法。方法针对马尔堡病毒、扎伊尔埃博拉病毒的特异性基因序列设计2对引物和相应的TSPE引物。经多重PCR扩增之后、加入连有TAG的TSPE引物特异性识别靶目标,标记有生物素的dCTP加入到延伸序列中,TAG与微球上的anti-TAG互补,SAPE与生物素反应,SAPE发射荧光信号,信号由液相芯片仪器检测。结果液相芯片检测体系能够正确的鉴定和检测两种出血热病毒,特异性强、灵敏度高,可用于病毒的高通量筛查。结论建立了多重PCR基因液相芯片快速检测两种出血热病毒的新方法。Objective To develop novel liquiehip array for detection of Marburg virus and Ebola virus. Methods Two pairs specific primer sets and multiplex Target-Specific Primer Extension primer were designed to amplify unique gene regions of Marburg virus and Ebola virus , Multiplex PCR amplification was carried by Biotin-dCTP, PCR products were hybridized to corresponding probe sequences coupling on the unique sets of beads, fluorescent signal of SAPE which reactive with Biotin was collected by Bio-plex workstation. Results Liquidchip array has good sensitivity and specificity for detection of Marburg virus and Ebola virus, Which has good prospects of application in high thoughput screening of virus. Conclusion The multiplex PCR liquidchip array was established for simultaneous detection of Marburg virus and Ebola virus.

关 键 词：液相芯片 马尔堡病毒 埃博拉病毒 多重PCR TSPE引物 

分 类 号：R512.8[医药卫生—内科学]