液相芯片检测3种发热伴出疹病毒方法的建立

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

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。

Objective To develop a rapid,high-throughput screening method of gene liquid chip to detect three viruses of fever and rash syndromes,including EV71,CA16 and Measles virus.Methods Highly validated specific primers and probes were used to amplify the target regions of each virus.Biotin labeled PCR products were hybridized to corresponding probes coupling on the unique fluorescent beads.The hybridized beads were processed through the Bio-plex,which identified the presence of PCR products.The sensitivity,specificity and detection power were also analyzed.Results The result of liquid chip on the detection of three kinds of viruses was consistent with the result of multiple PCR.The assay can detect each virus and all three kinds of viruses simultaneously,with high specificity and the lowest detection limitation was 2.14 pg/μl.Conclusion The assay,which is high sensitivity and specificity,can be used to detect EV71,CA16 and MV in the laboratory.