Tu Yunping, Yang Dianlong, Zhang Zhongping, Dong Xiaobin, Liu Luyao, Miao Guijun, Zhang Lulu, Qiu Xianbo
College of Information Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Mar 25;38(3):943-960. doi: 10.13345/j.cjb.210533.
Polymerase chain reaction (PCR) is the gold standard for nucleic acid amplification in molecular diagnostics. The PCR includes multiple reaction stages (denaturation, annealing, and extension), and a complicated thermalcycler is required to repetitively provide different temperatures for different stages for 30-40 cycles within at least 1-2 hours. Due to the complicated devices and the long amplification time, it is difficult to adopt conventional PCR in point-of-care testing (POCT). Comparing to conventional PCR, isothermal amplification is able to provide a much faster and more convenient nucleic acid detection because of highly efficient amplification at a constant reaction temperature provided by a simple heating device. When isothermal amplification is combined with microfluidics, a more competent platform for POCT can be established. For example, various diagnosis devices based on isothermal amplification have been used to rapidly and conveniently detect SARS-CoV-2 viruses. This review summarized the recent development and applications of the microfluidics-based isothermal amplification. First, different typical isothermal amplification methods and related detection methods have been introduced. Subsequently, different types of microfluidic systems with isothermal amplification were discussed based on their characteristics, for example, functionality, system structure, flow control, and operation principles. Furthermore, detection of pathogens (e.g. SARS-CoV-2 viruses) based on isothermal amplification was introduced. Finally, the combination of isothermal amplification with other new technologies, e.g. CRISPR, has been introduced as well.
聚合酶链反应（PCR）是分子诊断中核酸扩增的金标准。PCR包括多个反应阶段（变性、退火和延伸），并且需要一个复杂的热循环仪在至少1-2小时内为不同阶段重复提供不同温度达30-40个循环。由于设备复杂且扩增时间长，难以在即时检测（POCT）中采用传统PCR。与传统PCR相比，等温扩增能够在由简单加热装置提供的恒定反应温度下进行高效扩增，从而提供更快、更便捷的核酸检测。当等温扩增与微流控技术相结合时，可以建立一个更具竞争力的POCT平台。例如，基于等温扩增的各种诊断设备已被用于快速、方便地检测SARS-CoV-2病毒。本综述总结了基于微流控的等温扩增的最新发展和应用。首先，介绍了不同的典型等温扩增方法和相关检测方法。随后，根据其特点，如功能、系统结构、流量控制和操作原理，讨论了具有等温扩增功能的不同类型微流控系统。此外，还介绍了基于等温扩增的病原体（如SARS-CoV-2病毒）检测。最后，也介绍了等温扩增与其他新技术（如CRISPR）的结合。
