埃可病毒9型荧光RT-PCR快速检测方法的建立及应用

发布时间：2018-02-05 23:52

本文关键词： 埃可病毒型(ECHO-) 荧光定量RT-PCR TaqMan探针　出处：《病毒学报》2017年06期 　论文类型：期刊论文

【摘要】：为了建立特异、灵敏、快速的荧光定量RT-PCR方法用于检测埃可病毒9型病毒核酸,并初步应用于埃可病毒9型的临床标本检测。根据GenBank登录的埃可病毒9型毒株VP1基因序列,应用生物学软件进行序列比对,在保守区设计特异性引物和TaqMan探针。对反应条件进行优化,验证方法的特异性、灵敏度和重复性,同时对疑似埃可病毒9型病例标本进行检测。该方法对埃可病毒9型的检出有高度特异性,与EV71、CA16、CA24v、CA6、CA10、埃可病毒30型等均无交叉反应,检测灵敏度均达0.1TCID50/mL,可从疑似埃可病例粪便标本中直接检测病毒核酸,检测仅需3~4h。本研究建立的埃可病毒9型TaqMan荧光定量RT-PCR检测方法特异、灵敏,适用于临床早期诊断。
[Abstract]:In order to establish a specific, sensitive and rapid fluorescent quantitative RT-PCR method for the detection of the nucleic acid of Ecovirus type 9 virus. According to the VP1 gene sequence of EKV 9 strain registered by GenBank, the sequence alignment was carried out by using biological software. Specific primers and TaqMan probes were designed in the conserved region. The reaction conditions were optimized to verify the specificity, sensitivity and repeatability of the method. At the same time, the specimens of suspected EKV-9 cases were detected. The method was highly specific to EV71CA16C16 and CA24vCA6CA10. There was no cross reaction of EKV type 30 and the detection sensitivity was 0. 1 TCID 50 / mL, which could be directly detected from fecal specimens of suspected Eco cases. The method developed in this study is specific, sensitive and suitable for early clinical diagnosis of EKV type 9 TaqMan.
【作者单位】：杭州市疾病预防控制中心;
【基金】：杭州市科技发展计划项目(项目号:20120633B22);题目:埃可病毒9型的快速检测方法建立以及流行特征研究杭州市农业与社会发展科研计划项目(项目号:20170533B76);题目:杭州市区手足口病患儿中埃可病毒的检测及其分子特征研究~~
【分类号】：R440;R512.5

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