Paper-based nucleic acid testing techniques are increasingly in demand. Hence, we have developed a simple and cheap paper fluidic device to detect multiple gene targets in Leptospira. Fluidic channels of the penta-clover device are drawn using a correction pen on Whatman filter paper 1. The fluid blocks the pores of the paper, avoiding leakage and ensuring the equal flow of sample to the reaction pads. The target genes are amplified by performing Loop-Mediated Isothermal Amplification (LAMP) with dry reaction components. Thecolor change of leuco crystal violetallows real-time monitoring of a positive amplification. The difference in color intensity is captured with a smartphone and analyzed using image processing software. The device amplifies the target within 15 min, detects the pathogen at a concentration as low as 50 attogram μl⁻¹, detects Leptospira in blood samples without prior treatment and differentiates the Leptospira species even after 21 days of storage at room temperature.

纸基核酸测试技术的需求日益增长。因此，我们开发了一种简单廉价的纸质流体装置来检测钩端螺旋体中的多个基因靶标。使用校正笔在Whatman滤纸1上绘制五叶草设备的流体通道。流体会阻塞纸的孔，避免泄漏，并确保样品均匀流入反应垫。通过对干反应组分进行环介导的等温扩增（LAMP），可以扩增靶基因。无色晶体紫罗兰色的颜色变化可实时监测阳性扩增。用智能手机捕获颜色强度的差异，并使用图像处理软件进行分析。该设备可在15分钟内扩增靶标，检测浓度低至50 attogramμl ^-1的病原体，无需事先治疗即可检测血样中的钩端螺旋体，并区分钩端螺旋体 种即使在室温下保存21天也是如此。