Liao Jiaqi, Li Chonghao, Liu Jiajia, Cheng Hao, Li Linmin, Tang Guixi, Huang Ru, Lu Yating, Chen Siyue, Zhang Qian, Chen Hao, Chen Qinyuan, Chen Hong, Sun Daoheng
Pen-Tung Sah Institute of Micro-Nano Science and Technology, State Key Laboratory of Vaccines for Infectious Diseases, Xiang an Biomedicine Laboratory, Xiamen University, Xiamen, Fujian, 361005, China.
College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian, 361005, China.
Anal Chim Acta. 2025 Jan 15;1334:343415. doi: 10.1016/j.aca.2024.343415. Epub 2024 Nov 13.
The detection of nucleic acids, which typically consists of nucleic acid extraction, reverse transcription and amplification, is a crucial component of molecular diagnostics. However, traditional methods have several limitations, including high reliance on skilled personnel, low degree of automation and lengthy assay times. Additionally, these methods also require separate areas for the different steps to avoid contamination. Though a number of digital microfluidic (DMF) chips have been reported to detect the nucleic acids, but there is a noticeable absence of fully integrated DMF platforms capable of concurrently performing nucleic acid extraction, amplification, and signal detection. Here, a DMF chip combined with a dual-mode thermal control module was developed to realize the automatic and integrated detection of nucleic acids. The dual-mode thermal control module provided the required temperatures (with a range from 4 °C to 95 °C) for all the steps of nucleic acid detection. Pseudoviruses of SARS-CoV-2 and monkeypox were chosen to demonstrate the feasibility of the DMF chip, and all assays were completed within 40 min. The results confirmed that the products extracted under 4 °C exhibited high concentration and integrity, which were comparable to those by traditional methods. And the analytical performance of amplify was comparable to that of traditional methods with a more compact design, shorter time, smaller size and more automatic operation. With the ability of dual-mode thermo control and full automatic operation, nucleic acid detection with the characteristics of sample-in-result-out was realized on the DMF chip, which holds great promise in fields such as in vitro diagnostics and point-of-care testing.
核酸检测通常包括核酸提取、逆转录和扩增，是分子诊断的关键组成部分。然而，传统方法存在一些局限性，包括高度依赖技术人员、自动化程度低和检测时间长。此外，这些方法还需要为不同步骤设置单独的区域以避免污染。尽管已经报道了许多用于检测核酸的数字微流控（DMF）芯片，但明显缺乏能够同时进行核酸提取、扩增和信号检测的完全集成的DMF平台。在此，开发了一种结合双模式热控模块的DMF芯片，以实现核酸的自动和集成检测。双模式热控模块为核酸检测的所有步骤提供所需的温度（范围为4℃至95℃）。选择了新型冠状病毒2和猴痘的假病毒来证明DMF芯片的可行性，所有检测均在40分钟内完成。结果证实，在4℃下提取的产物具有高浓度和完整性，与传统方法相当。并且扩增的分析性能与传统方法相当，具有更紧凑的设计、更短的时间、更小的尺寸和更自动化的操作。凭借双模式热控和全自动操作能力，在DMF芯片上实现了具有样本进结果出特点的核酸检测，这在体外诊断和即时检测等领域具有广阔前景。
