Xu Ruiting, Saiduzzaman S M, Sampad Mohammad Julker Neyen, Wells Tanner, Cavinatto Isadora H, Bellido Edward M, Holliday Susannah, Ong Ephraim, Mdaki Stephanie, Tamhankar Manasi, Yuzvinsky Thomas D, Patterson Jean L, Hawkins Aaron, Schmidt Holger
School of Engineering, University of California, Santa Cruz, California 95064, United States.
Electrical and Computer Engineering Department, Brigham Young University, Provo, Utah 84602, United States.
ACS Sens. 2025 Aug 22;10(8):5772-5780. doi: 10.1021/acssensors.5c00926. Epub 2025 Aug 12.
Molecular diagnostics of biomarkers derived from tissues and bodily fluids are developing into a cornerstone of medical testing and potentially the foundation of a personalized medicine approach. Therefore, there is a need for universal biosensors capable of detecting different biomarker types with high performance and minimal complexity, ideally suitable for point-of-care use. Here, we present an integrated nanopore sensor for amplification-free, label-free, and quantitative detection of both proteins and nucleic acids from clinical samples. High performance is enabled by trapping target-carrying microbeads at the nanopore location and rapid detection of individual targets upon release from the beads. Low complexity is achieved via a simple solid-phase extraction protocol and optics-free, flow-based bead trapping. The device is validated through the analysis of biofluids from a baboon infected with SARS-CoV-2. Detection of the spike protein down to 100 fg/mL, with a linear count over 5 orders of magnitude, and SARS-CoV-2 RNA over the entire course of infection, down to a concentration of 7.5 aM, are demonstrated. This platform enables ultrasensitive, quantitative analysis of different biomarker types using a single protocol and holds promise as a universal biosensor for infectious disease detection and a wide range of applications.
源自组织和体液的生物标志物的分子诊断正发展成为医学检测的基石，并有可能成为个性化医疗方法的基础。因此，需要一种通用的生物传感器，能够以高性能和最小的复杂性检测不同类型的生物标志物，理想情况下适用于即时检测。在此，我们展示了一种集成纳米孔传感器，用于对临床样本中的蛋白质和核酸进行无扩增、无标记的定量检测。通过将携带靶标的微珠捕获在纳米孔位置，并在微珠释放后快速检测单个靶标，实现了高性能。通过简单的固相萃取方案以及无光学元件、基于流动的微珠捕获实现了低复杂性。该设备通过分析感染了SARS-CoV-2的狒狒的生物流体进行了验证。结果表明，能够检测低至100 fg/mL的刺突蛋白，线性计数超过5个数量级，并且能够在整个感染过程中检测低至7.5 aM浓度的SARS-CoV-2 RNA。该平台能够使用单一方案对不同类型的生物标志物进行超灵敏、定量分析，并有望成为用于传染病检测及广泛应用的通用生物传感器。
