两种出血热病毒新型液相芯片检测体系的建立

    杨洪1，2，王静1，刘衡川2，杨宇1，杨永莉1，张乐1

    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

    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。

    关键词：液相芯片；马尔堡病毒；埃博拉病毒；多重PCR；TSPE引物

    中图分类号：R512.8　文献标识码：B

    Novel liquidchip array of detection for twokinds of hemorrhagic fever virus

    YANG Hong*，WANG Jing，LIU Heng-chuan, YANG Yu,YANG Yong-li, ZHANG Le

    China Academy of Inspection and Quarantine ,Beijing 100025, China

    Abstract:   Objective  To develop novel liquichip array for detection of Marburg virus andEbola virus.   Methods   Two pairs specificprimer sets and multiplex Target-Specific Primer Extension primerwere designed to amplify unique gene regions of Marburg virus andEbola virus , Multiplex PCR amplification was carried byBiotin-dCTP , PCR products were hybridized to corresponding probesequences coupling on the unique sets of beads. fluorescent signalof SAPE which reactive with Biotin was collected by Bio-plexworkstation.  Results   Liquidchip array has goodsensitivity and specificity for detection of Marburg virus andEbola virus, Which has good prospects of application in highthoughput screening of virus.  Conclusion   Themultiplex PCR liquidchip array was established for simultaneousdetection of Marburg virus and Ebola virus.

    Key words:   Liquidchip array;Marburg virus; Ebola virus; Multiplex PCR; TSPE primer

    《中国国境卫生检疫杂志》