Department of Central Laboratory, Clinical Medicine Scientific and Technical Innovation Park, Shanghai Tenth People's Hospital, Shanghai, 200435, China.
Department of Clinical Laboratory, Yangpu Hospital, Tongji University School of Medicine, Shanghai, 200090, China.
Mikrochim Acta. 2022 May 4;189(5):212. doi: 10.1007/s00604-022-05283-9.
Loop-mediated isothermal amplification (LAMP) is a promising diagnostic tool for genetic amplification, which is known for its rapid process, simple operation, high amplification efficiency, and excellent sensitivity. However, most of the existing heating methods are external for completion of molecular amplification with possibility of contamination of specimens. The present research provided an internal heating method for LAMP using magnetic nanoparticles (MNPs), which is called nano-LAMP. Near-infrared light with an excitation wavelength of 808 nm was employed as the heating source; hydroxy naphthol blue (HNB) was used as an indicator to conduct methodological research. We demonstrate that the best temperature was controlled at a working power of 2 W and 4.8 µg/µL concentration of nanoparticles. The lowest limit for the detection of HPV by the nano-LAMP method is 10 copies/mL, which was confirmed by a gel electrophoresis assay. In the feasibility investigation of validated clinical samples, all 10 positive HPV-6 specimens amplified by nano-LAMP were consistent with conventional LAMP methods. Therefore, the nano-LAMP detection method using internal heating of MNPs may bring a new vision to the exploration of thermostatic detection in the future.
环介导等温扩增（LAMP）是一种很有前途的基因扩增诊断工具，其以快速、操作简单、扩增效率高、灵敏度高而著称。然而，大多数现有的加热方法都是外部的，用于完成分子扩增，存在标本污染的可能性。本研究采用磁性纳米粒子（MNPs）作为内部热源，提供了一种 LAMP 的内部加热方法，称为纳米 LAMP。近红外光（激发波长 808nm）作为加热源；羟基萘酚蓝（HNB）作为指示剂进行方法学研究。我们证明，最佳温度控制在工作功率为 2W 和纳米粒子浓度为 4.8μg/μL 时。纳米 LAMP 方法检测 HPV 的最低检测限为 10 拷贝/mL，凝胶电泳分析证实了这一点。在经过验证的临床样本的可行性研究中，纳米 LAMP 扩增的 10 份 HPV-6 阳性标本均与常规 LAMP 方法一致。因此，采用 MNPs 内部加热的纳米 LAMP 检测方法可能为未来恒温检测的探索带来新的视角。
