文章摘要
唐 震,李 显,郭喜玲,吴 涛,史智扬.一种基于NASBA技术的新型快速肠道病毒检测方法的应用研究[J].南京医科大学学报,2010,(12):1729~1731
一种基于NASBA技术的新型快速肠道病毒检测方法的应用研究
Development of a new method based on NASBA for enterovirus fast detection
投稿时间:2010-07-05  
DOI:10.7655
中文关键词: NASBA  磁珠  捕获探针  肠道病毒
英文关键词: NASBA, immunomagnetic beads, capture probes, enterovirus
基金项目:
作者单位
唐 震 江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,江苏 南京 210009 
李 显 江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,江苏 南京 210009 
郭喜玲 江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,江苏 南京 210009 
吴 涛 江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,江苏 南京 210009 
史智扬 江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,江苏 南京 210009 
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中文摘要:
      目的: 基于核酸等温扩增技术(nucleic acid sequence-based amplification,NASBA)荧光分子信标探针检测技术,进行肠道病毒的快速检测与辅助诊断?方法:根据Genebank上肠道病毒5′端非编码区序列,设计特异性引物与捕获探针,应用纳米技术对商品化磁珠进行偶联,NASBA方法进行扩增,NucliSens读数仪检测,荧光定量RT-PCR方法进行验证,同时验证NASBA方法的特异性?灵敏度和重复性?结果:该方法对肠道病毒具有高度特异性,与RSV等呼吸道相关病毒均无交叉反应,反应体系具有很高的稳定性?结论:本研究应用的肠道病毒NASBA检测方法特异?灵敏?快速简便,适用于肠道病毒的日常监测和爆发疫情的应急诊断?
英文摘要:
      Objective: To develop a new nucleic acid sequence-based amplification (NASBA) method which is based on fluorescence probe for the detection of enterovirus and clinic diagnosis. Methods: The special primers and probes were designed based on the nucleic acid sequences of relative enterovirus in Gene bank. The commercial immunomagectic beads were conjugated with the primers and probes using nanotechnology. The conjugated products were amplified by NASBA and sequentially detected by NucliSens reader. The results were validated by real time-PCR. The sensitivity, repetition and specificity of the method were evaluated respectively. Results: The method could detect enterovirus with high specificity and stability as well as no interactions with other respiratory virus such as RSV. Conclusion: The new NASBA method is convenience, fast, sensitive and specific for enterovirus detection, which could be applied for regular surveillance and urgent investigation of outbreaks of enterovirus in the near future.
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