Wang Kangning, Wang Qingran, Peng Canfu, Guo Yu, Li Yan, Zhou Jia, Wu Wenming
Institute of Biological and Medical Engineering, Guangdong Academy of Sciences, Guangzhou 516001, China.
State Key Laboratory of Luminescence and Applications, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun 130033, China.
ACS Omega. 2022 Jul 20;7(30):26165-26173. doi: 10.1021/acsomega.2c01824. eCollection 2022 Aug 2.
With the outbreak of COVID-19 around the world, rapid and accurate detection of new coronaviruses is the key to stop the transmission of the disease and prevent and control the novel coronavirus, among which polymerase chain reaction (PCR) is the mainstream nucleic acid detection method. A temperature cycling device is the core of the PCR amplification micro-device. The precision of the temperature control and temperature change rate directly affect the efficiency of PCR amplification. This study proposes a new PCR method based on rapid PCR chip optimization of a liquid metal bath, which realizes precise and rapid temperature rise and fall control. We systematically explored the feasibility of using liquid metals with different melting points in the system and proposed a 47 °C bismuth-based liquid metal bath as the heat conduction medium of the system to optimize the system. The heat conduction properties of the thermally conductive silicone oil bath were compared. Compared with the thermally conductive silicone oil bath, thermal cycle efficiency is improved nearly 3 times. The average heating rate of the liquid metal bath is fast, and the temperature control stability is good, which can significantly reduce the hysteresis, and the temperature change curve is more gentle, which can greatly improve the efficiency of PCR amplification. The results of gene amplification using rat DNA as the template and SEC61A as the target also indicate that the system can be successfully used in PCR devices, and the types of PCR containers can be not limited to PCR tubes. Based on the experiment, we proved that the PCR method optimized by the liquid metal bath multi-gene rapid PCR chip can further improve the temperature response speed. It has the advantages of accurate data, fast response speed, low price, safety, and environmental protection and can effectively reduce the time of PCR and improve the application efficiency. As far as we know, this is the first international report on using a liquid metal bath to do rapid-cooling PCR.
随着新冠疫情在全球爆发,快速准确地检测新型冠状病毒是阻断疾病传播、防控新型冠状病毒的关键,其中聚合酶链反应(PCR)是主流的核酸检测方法。温度循环装置是PCR扩增微装置的核心,温度控制的精度和温度变化速率直接影响PCR扩增效率。本研究提出一种基于液态金属浴快速PCR芯片优化的新型PCR方法,实现了精确快速的升降温控制。我们系统地探索了在该体系中使用不同熔点液态金属的可行性,并提出以47℃铋基液态金属浴作为体系的热传导介质来优化该体系。对比了导热硅油浴的热传导性能。与导热硅油浴相比,热循环效率提高了近3倍。液态金属浴的平均升温速率快,温度控制稳定性好,能显著降低滞后现象,温度变化曲线更平缓,可大大提高PCR扩增效率。以大鼠DNA为模板、SEC61A为靶点进行基因扩增的结果也表明,该体系可成功应用于PCR装置,PCR容器类型可不局限于PCR管。基于实验,我们证明了液态金属浴多基因快速PCR芯片优化的PCR方法可进一步提高温度响应速度。其具有数据准确、响应速度快、价格低廉、安全环保等优点,能有效缩短PCR时间,提高应用效率。据我们所知,这是关于使用液态金属浴进行快速冷却PCR的首份国际报告。