Kim Sungwan, Cho Giwon, Lee Jaebaek, Doshi Khushi, Gharpure Supriya, Kim Jisan, Gwak Juyong, Hardie Joseph M, Kanakasabapathy Manoj K, Kandula Hemanth, Thirumalaraju Prudhvi, Song Younseong, Chen Hui, Kuritzkes Daniel R, Li Jonathan Z, Tsibris Athe M, Shafiee Hadi
Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Nat Biomed Eng. 2025 May 30. doi: 10.1038/s41551-025-01405-9.
Bioluminescence holds notable promise as a modality in diagnostics due to its high signal-to-noise ratio and absence of incident radiation. However, challenges arise from rapid signal decay and reduced enzyme activity when linked to targeting molecules, limiting its reliability in point-of-care diagnostic applications. Here we introduce the luminescence cascade-based sensor (LUCAS) assay, an enzyme cascade system capable of detecting analytes with ultrahigh sensitivity and prolonged bioluminescence. Utilizing a sequential enzymatic reaction, our assay achieves a greater than 500-fold increase in bioluminescence signal and maintains an 8-fold improvement in signal persistence compared to conventional bioluminescence assays. Implemented on a portable, fully automated device designed for point-of-care settings, our system facilitates rapid (<23 min) sample-to-answer analysis of viruses without an external power supply. Its accuracy surpasses 94% in the qualitative classification of 177 viral-infected patient samples and 130 viral-spiked serum samples, various pathogens including the respiratory virus SARS-CoV-2, and blood-borne pathogens such as HIV, HBV and HCV as clinical models. The decentralized, rapid, sensitive, specific and cost-effective nature of LUCAS positions it as a viable diagnostic tool for low-resource environments.
由于生物发光具有高信噪比且无入射辐射，它在诊断领域具有显著的应用前景。然而，当与靶向分子连接时，快速的信号衰减和酶活性降低会带来挑战，限制了其在即时诊断应用中的可靠性。在此，我们介绍基于发光级联的传感器（LUCAS）检测法，这是一种能够以超高灵敏度和延长生物发光来检测分析物的酶级联系统。利用顺序酶促反应，我们的检测法实现了生物发光信号增加超过500倍，与传统生物发光检测法相比，信号持久性提高了8倍。在专为即时检测环境设计的便携式全自动设备上实施，我们的系统无需外部电源即可快速（<23分钟）完成病毒样本到结果的分析。在以包括呼吸道病毒SARS-CoV-2以及血液传播病原体如HIV、HBV和HCV等各种病原体作为临床模型的177份病毒感染患者样本和130份病毒加标血清样本的定性分类中，其准确率超过94%。LUCAS分散、快速、灵敏、特异且经济高效的特性使其成为低资源环境下可行的诊断工具。
