Center for Soft and Living Matter, Institute for Basic Science (IBS), Ulsan 44919, Republic of Korea.
Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
Acc Chem Res. 2021 Oct 5;54(19):3643-3655. doi: 10.1021/acs.accounts.1c00367. Epub 2021 Sep 13.
Reliable, inexpensive, and rapid diagnostic tools are essential to control and prevent the spread of infectious diseases. Many commercial kits for coronavirus disease 2019 (COVID-19) diagnostics have played a crucial role in the fight against the COVID-19 pandemic. Most current standard diagnostic (IVD) protocols for infectious diseases are sensitive but time-consuming and require sophisticated laboratory equipment and specially trained personnel. Recent advances in biosensor technology suggest the potential to deliver point-of-care (POC) diagnostics that are affordable and provide accurate results in a short time. The ideal "sample-in-answer-out" type fully integrated POC infection diagnostic platforms are expected to be autonomous or easy-to-operate, equipment-free or infrastructure-independent, and high-throughput or easy to upscale. In this Account, we detail the recent progress made by our group and others in the development of centrifugal microfluidic devices or lab-on-a-disc (LOAD) systems. Unlike conventional pump-based fluid actuation, the centrifugal force generated by spinning the disc induces liquid pumping and no external fluidic interconnects are required. This allows a total fluidic network required for multiple steps of biological assays to be integrated on a disc, enabling fully automated POC diagnostics. Various applications have been demonstrated, including liquid biopsy for personalized cancer management, food applications, and environmental monitoring; here, we focus on IVD for infectious disease. First, we introduce various on-disc unit operation technologies, including reagent storage, sedimentation, filtration, valving, decanting, aliquoting, mixing, separation, serial dilution, washing, and calibration. Such centrifugal microfluidic technologies have already proved promising for micro-total-analysis systems for automated IVD ranging from molecular detection of pathogens to multiplexed enzyme-linked immunosorbent assays (ELISAs) that use raw samples such as whole blood or saliva. Some recent examples of LOAD systems for molecular diagnostics in which some or all steps of the assays are integrated on a disc, including pathogen enrichment, nucleic acid extraction, amplification, and detection, are discussed in detail. We then introduce fully automated ELISA systems with enhanced sensitivity. Furthermore, we demonstrate a toy-inspired fidget spinner that enables electricity-free and rapid analysis of pathogens from undiluted urine samples of patients with urinary tract infection symptoms and a phenotypic antimicrobial susceptibility test for an extreme POC diagnostics application. Considering the urgent need for cost-effective and reliable POC infection diagnostic tools, especially in the current pandemic crisis, the current limitations and future directions of fast and broad adaptation in real-world settings are also discussed. With proper attention to key challenges and leverage with recent advances in bio-sensing technologies, molecular biology, nanomaterials, analytical chemistry, miniaturization, system integration, and data management, LOAD systems hold the potential to deliver POC infection diagnostic tools with unprecedented performance regarding time, accuracy, and cost. We hope the new insight and promise of LOAD systems for POC infection diagnostics presented in this Account can spark new ideas and inspire further research and development to create better healthcare systems for current and future pandemics.
可靠、廉价且快速的诊断工具对于控制和预防传染病的传播至关重要。许多用于 2019 年冠状病毒病(COVID-19)诊断的商业试剂盒在抗击 COVID-19 大流行方面发挥了关键作用。大多数当前用于传染病的标准诊断(IVD)方案虽然灵敏,但耗时且需要复杂的实验室设备和经过专门培训的人员。最近生物传感器技术的进步表明,有可能提供负担得起的即时准确结果的即时护理(POC)诊断。理想的“样本进答案出”型完全集成的 POC 感染诊断平台预计是自主或易于操作的,无设备或独立于基础设施的,高通量或易于扩展的。在本报告中,我们详细介绍了我们小组和其他小组在开发离心微流体装置或盘上实验室(LOAD)系统方面取得的最新进展。与传统的基于泵的流体致动不同,通过旋转盘产生的离心力诱导液体泵送,并且不需要外部流体互连。这允许将用于生物测定的多个步骤的总流体网络集成在一个盘上,从而实现全自动 POC 诊断。已经证明了各种应用,包括用于个性化癌症管理的液体活检、食品应用和环境监测;在这里,我们专注于传染病的 IVD。首先,我们介绍了各种盘上单元操作技术,包括试剂储存、沉淀、过滤、阀、倾析、分注、混合、分离、连续稀释、洗涤和校准。这种离心微流体技术已经证明对于用于自动化 IVD 的微全分析系统很有前途,从病原体的分子检测到使用全血或唾液等原始样本的多重酶联免疫吸附测定(ELISA)。详细讨论了一些最近的 LOAD 系统用于分子诊断的示例,其中一些或所有测定步骤都集成在一个盘上,包括病原体富集、核酸提取、扩增和检测。然后,我们介绍了具有增强灵敏度的全自动 ELISA 系统。此外,我们展示了一个受玩具启发的指尖陀螺,它可以实现无需电源的快速分析,用于从有尿路感染症状的患者的未稀释尿液样本中分离病原体,并进行针对极端 POC 诊断应用的表型抗菌药物敏感性测试。考虑到对经济实惠且可靠的 POC 感染诊断工具的迫切需求,特别是在当前的大流行危机中,还讨论了在现实环境中快速广泛适应的当前限制和未来方向。通过适当关注关键挑战,并利用生物传感技术、分子生物学、纳米材料、分析化学、小型化、系统集成和数据管理方面的最新进展,LOAD 系统有可能提供具有前所未有的时间、准确性和成本性能的 POC 感染诊断工具。我们希望本报告中呈现的用于 POC 感染诊断的 LOAD 系统的新见解和前景能够激发新的想法,并激励进一步的研究和开发,为当前和未来的大流行创造更好的医疗保健系统。
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