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

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

吉林大学药学院, 长春 130021

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。

关键词:

English

Detection of Cytomegalovirus Based on Loop-Mediated Isothermal Amplification Combined with Surface Enhanced Raman Spectroscopy

School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China

Corresponding author: LIANG Chong-Yang, liang@jlu.edu.cn

Received Date: 04 January 2022
Revised Date: 18 January 2022
Fund Project:
Supported by the Science and Technology Planning Project of Jilin Province, China (No.20190304035YY).

Abstract: Cytomegalovirus (CMV) is very harmful to patients with organ transplantation, pregnant women and newborns. At present,quantitative real-time PCR (qPCR) is the most important method for clinical detection of CMV. However, due to the high cost and complex operation of qPCR, it is difficult to be applied in rapid detection on site. Loop mediated isothermal amplification (LAMP) is a sensitive, simple and rapid nucleic acid detection technology. LAMP shows better amplification efficiency than qPCR, but it is easy to produce dimer between multiple pairs of primers, which will result in nonspecific amplification, and thus cause false positive results. In this study, two probes with complementary sequences were designed to connect magnetic beads (MNs) and silver nanoparticles (AgNPs) modified with Raman probe signal molecule 4-mercaptobenzoic acid (4-MBA). A stronger signal on the surface of AgNPs was obtained by surface enhanced Raman scattering. By using the designed method with the above principle, the standard nucleic acid sequence of CMV was detected, and the detection limit reached 1000 copies/mL. At the level of structural principle of amplification products, the false positive problem perplexing LAMP technology was solved, which showed reference value for the development of new rapid detection technology.

Key words:

Cytomegalovirus
/ Loop-mediated isothermal amplification
/ Surface enhanced Raman spectroscopy
/ 4-Mercaptobenzoic acid

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沈阳化工大学材料科学与工程学院沈阳 110142

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

English

Detection of Cytomegalovirus Based on Loop-Mediated Isothermal Amplification Combined with Surface Enhanced Raman Spectroscopy

Corresponding author: LIANG Chong-Yang, liang@jlu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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

English

Detection of Cytomegalovirus Based on Loop-Mediated Isothermal Amplification Combined with Surface Enhanced Raman Spectroscopy

Corresponding author: LIANG Chong-Yang, liang@jlu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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