High fluorescence quenching probe-based reverse fluorescence enhancement LFTS coupling with IS-primer amplification reaction for the rapid and sensitive Parkinson Disease-associated MicroRNA detection

被引:26
作者
Chen, Minghui [1 ,2 ]
Li, Hengxuan [1 ,2 ]
Shi, Zhihong [3 ]
Peng, Weipan [1 ,2 ]
Qin, Yi [1 ,2 ]
Luo, Ran [1 ,2 ]
Zhou, Dianming [4 ]
Gong, Xiaoqun [1 ,2 ]
Chang, Jin [1 ,2 ]
机构
[1] Tianjin Univ, Sch Life Sci, Tianjin 300072, Peoples R China
[2] Tianjin Engn Ctr Micronano Biomat & Detect Treatm, Tianjin 300072, Peoples R China
[3] Tianjin Huanhu Hosp, Dept Neurol, Tianjin 300350, Peoples R China
[4] Tianjin Ctr Dis Control & Prevent, Dept Toxicol, Tianjin 300011, Peoples R China
基金
中国国家自然科学基金;
关键词
Parkinson disease; miRNA; rLFTS; High fluorescence quenching probe; ISAR; LATERAL FLOW DEVICE; SERS DETECTION; BIOMARKERS; MIRNA; MECHANISM; CANCER; STRIP; RNA;
D O I
10.1016/j.bios.2020.112278
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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.
引用
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页数:8
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