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

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: 4 January 2022
Revised Date: 18 January 2022

Fund Project: Supported by the Science and Technology Planning Project of Jilin Province, China (No.20190304035YY).

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