MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou 510631, China; College of Biophotonics, South China Normal University, Guangzhou 510631, China; Department of Laboratory Medicine, Guangzhou First People's Hospital & The Second Affiliated Hospital of South China University of Technology, Guangzhou 510180, China.
MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou 510631, China; College of Biophotonics, South China Normal University, Guangzhou 510631, China.
Biosens Bioelectron. 2017 Nov 15;97:360-368. doi: 10.1016/j.bios.2017.06.014. Epub 2017 Jun 8.
Timely and accurate molecular diagnostics at the point-of-care (POC) level is critical to global health. To this end, we propose a handheld convective-flow real-time polymerase chain reaction (PCR) system capable of direct sample-to-answer genetic analysis for the first time. Such a system mainly consists of a magnetic bead-assisted photothermolysis sample preparation, a closed-loop convective PCR reactor, and a wireless video camera-based real-time fluorescence detection. The sample preparation exploits the dual functionality of vancomycin-modified magnetic beads (VMBs) for bacteria enrichment and photothermal conversion, enabling cell pre-concentration and lysis to be finished in less than 3min. On the presented system, convective thermocycling is driven by a single-heater thermal gradient, and its amplification is monitored in real-time, with an analysis speed of less than 25min, a dynamic linear range from 10 to 10 copies/µL and a detection sensitivity of as little as 1 copies/µL. Additionally, the proposed PCR system is self-contained with a control electronics, pocket-size and battery-powered, providing a low-cost genetic analysis in a portable format. Therefore, we believe that this integrated system may become a potential candidate for fast, accurate and affordable POC molecular diagnostics.
即时、准确的床边即时分子诊断对全球健康至关重要。为此,我们首次提出了一种手持式对流实时聚合酶链反应(PCR)系统,能够直接进行样本到答案的基因分析。该系统主要包括磁珠辅助光热解样品制备、闭环对流 PCR 反应器和基于无线摄像头的实时荧光检测。样品制备利用万古霉素修饰的磁珠(VMB)的双重功能进行细菌富集和光热转换,在不到 3 分钟的时间内完成细胞预浓缩和裂解。在本系统中,对流热循环由单个加热器热梯度驱动,并实时监测其扩增,分析速度小于 25 分钟,动态线性范围为 10 至 10 拷贝/μL,检测灵敏度低至 1 拷贝/μL。此外,所提出的 PCR 系统是一个独立的系统,具有控制电子设备,尺寸小巧,电池供电,以便携式格式提供低成本的遗传分析。因此,我们相信该集成系统可能成为快速、准确和经济实惠的床边即时分子诊断的潜在候选者。