Point-of-care testing using paper-based lateral flow assays (LFAs) has emerged as an attractive diagnostic platform. However, detecting small molecules such as cortisol using LFAs is challenging due to limited binding sites and weak signal generation. Here, we report the development of cortisol-specific aptamer-based probes and a paper-based origami preconcentrator (POP) to amplify the probe signal. The cortisol-specific aptamers were conjugated onto gold nanoparticles and hybridized with signal probes to create the cortisol-specific signal-on probe. POP, consisting of patterned layers with convergent wicking zones, induces electrokinetic preconcentration of the released signaling probes. By integrating cortisol-selective aptamer-based probes and POP, we accurately diagnosed cortisol levels within 30 min of signal probe incubation, followed by 10 min of preconcentration. Our sensor was able to detect cortisol levels in the range of 25–1000 ng/mL, with typical cortisol levels in plasma ranging from 40 to 250 ng/mL falling within this range. The successful detection of the wide range of cortisol samples using this approach highlights the potential of this platform as a point-of-care testing tool, particularly for lateral flow assay-based detection of small molecules like cortisol. Our approach offers a convenient and reliable method of cortisol level testing with a portable and accessible diagnosis device.

使用纸质侧流检测 (LFA) 的即时检测已成为一种有吸引力的诊断平台。然而，由于结合位点有限且信号生成较弱，使用 LFA 检测皮质醇等小分子具有挑战性。在这里，我们报告了皮质醇特异性适体探针和纸基折纸预浓缩器（POP）的开发，以放大探针信号。将皮质醇特异性适体缀合到金纳米粒子上并与信号探针杂交以产生皮质醇特异性信号探针。POP 由具有会聚芯吸区域的图案层组成，可诱导释放的信号探针的电动预浓缩。通过整合基于皮质醇选择性适体的探针和 POP，我们在信号探针孵育 30 分钟内准确诊断了皮质醇水平，然后预浓缩 10 分钟。我们的传感器能​​够检测 25–1000 ng/mL 范围内的皮质醇水平，血浆中典型的皮质醇水平范围为 40 至 250 ng/mL，属于该范围。使用这种方法成功检测各种皮质醇样品凸显了该平台作为即时检测工具的潜力，特别是基于侧流分析的皮质醇等小分子检测。我们的方法通过便携式且易于使用的诊断设备提供了一种方便可靠的皮质醇水平测试方法。使用这种方法成功检测各种皮质醇样品凸显了该平台作为即时检测工具的潜力，特别是基于侧流分析的皮质醇等小分子检测。我们的方法通过便携式且易于使用的诊断设备提供了一种方便可靠的皮质醇水平测试方法。使用这种方法成功检测各种皮质醇样品凸显了该平台作为即时检测工具的潜力，特别是基于侧流分析的皮质醇等小分子检测。我们的方法通过便携式且易于使用的诊断设备提供了一种方便可靠的皮质醇水平测试方法。