Nguyen Kim Truc, Rima Xilal Y, Nguyen Luong T H, Wang Xinyu, Kwak Kwang Joo, Yoon Min Jin, Li Hong, Chiang Chi-Ling, Doon-Ralls Jacob, Scherler Kelsey, Fallen Shannon, Godfrey Stephanie L, Wallick Julie A, Magaña Setty M, Palmer Andre F, Lee Inyoul, Nunn Christopher C, Reeves Kimberly M, Kaplan Henry G, Goldman Jason D, Heath James R, Wang Kai, Pancholi Preeti, Lee L James, Reátegui Eduardo
William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH, 43210, USA.
Diabetes and Metabolism Research Center, The Ohio State University Wexner Medical Center, Columbus, OH, 43210, USA.
Adv Healthc Mater. 2025 Jan;14(3):e2400622. doi: 10.1002/adhm.202400622. Epub 2024 Jun 23.
Virion-mediated outbreaks are imminent and despite rapid responses, continue to cause adverse symptoms and death. Therefore, tunable, sensitive, high-throughput assays are needed to help diagnose future virion-mediated outbreaks. Herein, it is developed a tunable in situ assay to selectively enrich virions and extracellular vesicles (EVs) and simultaneously detect antigens and nucleic acids at a single-particle resolution. The Biochip Antigen and RNA Assay (BARA) enhanced sensitivities compared to quantitative reverse-transcription polymerase chain reaction (qRT-PCR), enabling the detection of virions in asymptomatic patients, genetic mutations in single virions, and enabling the continued long-term expression of viral RNA in the EV-enriched subpopulation in the plasma of patients with post-acute sequelae of the coronavirus disease of 2019 (COVID-19). BARA revealed highly accurate diagnoses of COVID-19 by simultaneously detecting the spike glycoprotein and nucleocapsid-encoding RNA in saliva and nasopharyngeal swab samples. Altogether, the single-particle detection of antigens and viral RNA provides a tunable framework for the diagnosis, monitoring, and mutation screening of current and future outbreaks.
病毒体介导的疫情迫在眉睫，尽管采取了快速应对措施，但仍继续导致不良症状和死亡。因此，需要可调谐、灵敏、高通量的检测方法来帮助诊断未来由病毒体介导的疫情。在此，开发了一种可调谐原位检测方法，以选择性富集病毒体和细胞外囊泡（EVs），并在单颗粒分辨率下同时检测抗原和核酸。与定量逆转录聚合酶链反应（qRT-PCR）相比，生物芯片抗原和RNA检测（BARA）提高了灵敏度，能够检测无症状患者中的病毒体、单个病毒体中的基因突变，并能使2019冠状病毒病（COVID-19）急性后遗症患者血浆中富含EV的亚群中的病毒RNA持续长期表达。BARA通过同时检测唾液和鼻咽拭子样本中的刺突糖蛋白和编码核衣壳的RNA，实现了对COVID-19的高度准确诊断。总之，对抗原和病毒RNA的单颗粒检测为当前和未来疫情的诊断、监测和突变筛查提供了一个可调谐框架。
