基于环介导等温扩增联合表面增强拉曼光谱检测巨细胞病毒的研究

关鑫 武毅 徐冰 金正洋 梁重阳

引用本文: 关鑫, 武毅, 徐冰, 金正洋, 梁重阳. 基于环介导等温扩增联合表面增强拉曼光谱检测巨细胞病毒的研究[J]. 分析化学, 2022, 50(3): 424-432. doi: 10.19756/j.issn.0253-3820.221007 shu
Citation:  GUAN Xin,  WU Yi,  XU Bing,  JIN Zheng-Yang,  LIANG Chong-Yang. Detection of Cytomegalovirus Based on Loop-Mediated Isothermal Amplification Combined with Surface Enhanced Raman Spectroscopy[J]. Chinese Journal of Analytical Chemistry, 2022, 50(3): 424-432. doi: 10.19756/j.issn.0253-3820.221007 shu

基于环介导等温扩增联合表面增强拉曼光谱检测巨细胞病毒的研究

    通讯作者: 梁重阳,E-mail:liang@jlu.edu.cn
  • 基金项目:

    吉林省科技计划项目(No.20190304035YY)资助。

摘要: 巨细胞病毒(Cytomegalovirus,CMV)对器官移植患者、产妇及新生儿等免疫功能失调人群的危害极大。目前,临床检测CMV的主要方法为荧光定量PCR法(Quantitative polymerase chain reaction,qPCR),但其检测成本高,仪器操作复杂,不适于现场快速检测。环介导等温扩增(Loop-mediated isothermal amplification,LAMP)是一种超灵敏且简单快速的核酸检测技术,其扩增效率优于qPCR,但其引物间易产生二聚体,导致非特异性扩增,从而出现假阳性结果。本研究针对LAMP扩增产物中的DNA单链环状结构,设计了两段序列互补的探针,分别连接磁珠(Magnetic beads,MNs)和修饰有拉曼探针信号分子4-巯基苯甲酸(4-Mercaptobenzoic acid,4-MBA)的银纳米粒子(Sliver nanoparticles,AgNPs),二者与环状结构结合,形成MNs-LAMP产物-AgNPs复合物。通过磁场富集此复合物,表面增强拉曼散射(Surface enhanced Raman scattering,SERS)光谱监测结果表明,复合物的SERS信号更强。采用本方法检测了CMV标准核酸序列,检出限低至1000 copies/mL。本研究在扩增产物结构原理层面上解决了LAMP技术可能出现假阳性结果的问题,对研发新型快速核酸检测技术具有参考价值。

English


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  • 收稿日期:  2022-01-04
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