液相芯片检测3种发热伴出疹病毒方法的建立

    刘丽娟1，姜超2，杨永莉1，王莎莎2，于珊珊1，王旺1

    1.中国检验检疫科学研究院，北京 100123；2. 沈阳农业大学

    摘要：目的建立一种能同时检测发热伴出疹症候群中的肠道病毒71（EV71）、柯萨奇病毒A组16（CA16）和麻疹病毒（MV）的高通量液相芯片检测方法。方法针对EV71、CA16和MV全基因组序列，分别设计特异性引物和探针，经多重PCR扩增并用生物素标记相应的基因片段，建立EV71、CA16和MV高通量核酸液相芯片检测方法。并对该方法灵敏度、特异度进行验证。结果液相芯片检测方法对EV71、CA16和MV的检测结果与多重PCR的检测结果一致。该方法能同时完成对其中任何1种及3种病毒的检测，特异性好，灵敏度最低可达2.14pg/μl。结论该方法用于3种发热伴出疹症候群病毒的检测具有较高的灵敏度和特异性，可用于EV71、CA16和MV的实验室检测。

    关键词：液相芯片；EV71；CA16；MV；发热伴出疹

    中图分类号：R512.5 R511.1 R511.7 　文献标识码：B

    Development of a liquid chip for simultaneousdetection on three kinds of viruses of fever and rash syndromes

    LIU Li-juan*, JIANG Chao, YANG Yong-li, WANGSha-sha, YU Shan-shan, WANG Wang

    *Institute of Health Quarantine, ChineseAcademy of Inspeciton and Quarnatine, Beijing 100123,China

    Abstract:   Objective  To develop a rapid, high-throughput screening method of gene liquidchip to detect three viruses of fever and rash syndromes, includingEV71, CA16 and Measles virus.   Methods   Highly validated specific primers and probes were used to amplifythe target regions of each virus. Biotin labeled PCR products werehybridized to corresponding probes coupling on the uniquefluorescent beads. The hybridized beads were processed through theBio-plex, which identified the presence of PCR products. Thesensitivity, specificity and detection power were alsoanalyzed.   Results   The result of liquid chipon the detection of three kinds of viruses was consistent with theresult of multiple PCR. The assay can detect each virus and allthree kinds of viruses simultaneously, with high specificity andthe lowest detection limitation was 2.14 pg/μl. Conclusion    The assay, which is high sensitivityand specificity, can be used to detect EV71, CA16 and MV in thelaboratory.

    Key words:   Liquid chip; EV 71; CA16; MV; Fever and rash syndrome

    《中国国境卫生检疫杂志》2012年第5期