Parkinson Disease (PD) is the second-most common neurodegenerative disorder in the population. Recent researches indicated that hsa-microRNA 5010-3p (miR-5010) and hsa-microRNA 331-5p (miR-331) were significantly important for the detection of PD. So, in this work, a kind of high fluorescence quenching probe-based reverse fluorescence enhancement lateral flow test strip (rLFTS) was constructed to realize the synchronous detection of miR-5010 and miR-331. The formation of black hole quencher 2 (BHQ2) coating gold nanoparticles (AuNPs) effectively enhanced the fluorescence quenching property of the probes so as to significantly improve the detection sensitivity. This rLFTS also coupled with “invading stacking primer” (IS-primer) isothermal amplification reaction (ISAR) to accomplish rapid, sensitive, specific, and synchronous detection of PD-associated microRNA (miRNA). The whole detection time was shorter (35 min), and the limit-of-detection (LOD) reached to fM level. For the high accuracy diagnosis of PD, the synchronous determination of miR-5010 and miR-331 was successfully realized on one rLFTS by labeling fluorescent molecules to different T-line. This rLFTS also allowed for miRNA detection in total microRNA extracts from whole blood samples of PD patients, which performed important value in PD diagnosis and biomedical research.

帕金森病（PD）是人口中第二常见的神经退行性疾病。最近的研究表明，hsa-microRNA 5010-3p（miR-5010）和hsa-microRNA 331-5p（miR-331）对于检测PD至关重要。因此，在这项工作中，构建了一种基于高荧光猝灭探针的反向荧光增强侧流试纸（rLFTS），以实现miR-5010和miR-331的同步检测。黑洞淬灭剂2（BHQ2）涂层金纳米粒子（AuNPs）的形成有效增强了探针的荧光淬灭性能，从而大大提高了检测灵敏度。该rLFTS还可与“入侵堆叠引物”（IS引物）等温扩增反应（ISAR）结合使用，以实现快速，灵敏，特异的 和与PD相关的microRNA（miRNA）的同步检测。整个检测时间更短（35分钟），并且检测限（LOD）达到了fM水平。对于PD的高精度诊断，通过将荧光分子标记在不同的T线上，在一个rLFTS上成功实现了miR-5010和miR-331的同步测定。该rLFTS还可以检测PD患者全血样品中的总microRNA提取物中的miRNA，这在PD诊断和生物医学研究中具有重要价值。