Pumford Elizabeth A, Lu Jiakun, Spaczai Iza, Prasetyo Matthew E, Zheng Elaine M, Zhang Hanxu, Kamei Daniel T
Department of Bioengineering, Henry Samueli School of Engineering and Applied Sciences, University of California, Los Angeles, CA, 90095, USA.
Department of Bioengineering, Henry Samueli School of Engineering and Applied Sciences, University of California, Los Angeles, CA, 90095, USA.
Biosens Bioelectron. 2020 Dec 15;170:112674. doi: 10.1016/j.bios.2020.112674. Epub 2020 Oct 2.
Early disease detection through point-of-care (POC) testing is vital for quickly treating patients and preventing the spread of harmful pathogens. Disease diagnosis is generally accomplished using quantitative polymerase chain reaction (qPCR) to amplify nucleic acids in patient samples, permitting detection even at low target concentrations. However, qPCR requires expensive equipment, trained personnel, and significant time. These resources are not available in POC settings, driving researchers to instead utilize isothermal amplification, conducted at a single temperature, as an alternative. Common isothermal amplification methods include loop-mediated isothermal amplification, recombinase polymerase amplification, rolling circle amplification, nucleic acid sequence-based amplification, and helicase-dependent amplification. There has been a growing interest in combining such amplification methods with POC detection methods to enable the development of diagnostic tests that are well suited for resource-limited settings as well as developed countries performing mass screenings. Exciting developments have been made in the integration of these two research areas due to the significant impact that such approaches can have on healthcare. This review will primarily focus on advances made by North American research groups between 2015 and June 2020, and will emphasize integrated approaches that reduce user steps, reliance on expensive equipment, and the system's time-to-result.
通过即时检测(POC)进行疾病早期检测对于快速治疗患者和防止有害病原体传播至关重要。疾病诊断通常使用定量聚合酶链反应(qPCR)来扩增患者样本中的核酸,即使在低目标浓度下也能进行检测。然而,qPCR需要昂贵的设备、训练有素的人员,且耗时较长。这些资源在即时检测环境中无法获得,促使研究人员转而采用在单一温度下进行的等温扩增作为替代方法。常见的等温扩增方法包括环介导等温扩增、重组酶聚合酶扩增、滚环扩增、基于核酸序列的扩增和螺旋酶依赖性扩增。将此类扩增方法与即时检测方法相结合,以开发适用于资源有限环境以及进行大规模筛查的发达国家的诊断测试,这种兴趣日益浓厚。由于此类方法对医疗保健具有重大影响,这两个研究领域的整合取得了令人兴奋的进展。本综述将主要关注北美研究团队在2015年至2020年6月期间取得的进展,并将强调减少用户操作步骤、对昂贵设备的依赖以及系统出结果时间的整合方法。