Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Lembah Pantai, 50603 Kuala Lumpur, Malaysia.
The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, PR China and Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, PR China.
Analyst. 2016 May 10;141(10):2930-9. doi: 10.1039/c5an02532j.
Lateral flow assays (LFAs) have been extensively explored in nucleic acid testing (NAT) for medical diagnostics, food safety analysis and environmental monitoring. However, the amount of target nucleic acid in a raw sample is usually too low to be directly detected by LFAs, necessitating the process of amplification. Even though cost-effective paper-based amplification techniques have been introduced, they have always been separately performed from LFAs, hence increasing the risk of reagent loss and cross-contaminations. To date, integrating paper-based nucleic acid amplification into colorimetric LFA in a simple, portable and cost-effective manner has not been introduced. Herein, we developed an integrated LFA with the aid of a specially designed handheld battery-powered system for effective amplification and detection of targets in resource-poor settings. Interestingly, using the integrated paper-based loop-mediated isothermal amplification (LAMP)-LFA, we successfully performed highly sensitive and specific target detection, achieving a detection limit of as low as 3 × 10(3) copies of target DNA, which is comparable to the conventional tube-based LAMP-LFA in an unintegrated format. The device may serve in conjunction with a simple paper-based sample preparation to create a fully integrated paper-based sample-to-answer diagnostic device for point-of-care testing (POCT) in the near future.
侧向流动检测 (LFA) 在医学诊断、食品安全分析和环境监测等核酸检测 (NAT) 领域得到了广泛的探索。然而，原始样本中的目标核酸含量通常太低，无法直接通过 LFA 检测，因此需要进行扩增。尽管已经引入了具有成本效益的基于纸张的扩增技术，但它们始终与 LFAs 分开进行，因此增加了试剂损失和交叉污染的风险。迄今为止，尚未以简单、便携和具有成本效益的方式将基于纸张的核酸扩增集成到比色 LFA 中。在这里，我们开发了一种集成的 LFA，借助专门设计的手持电池供电系统，在资源匮乏的环境中有效扩增和检测目标。有趣的是，使用集成的基于纸张的环介导等温扩增 (LAMP)-LFA，我们成功地进行了高度敏感和特异性的目标检测，检测限低至 3×10(3) 个目标 DNA 拷贝，与非集成格式的传统基于管的 LAMP-LFA 相当。该设备可与简单的基于纸张的样品制备结合使用，以创建一种完全集成的基于纸张的样品到答案诊断设备，用于即时护理检测 (POCT) 在不久的将来。
