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A microfluidic paper-based laser-induced fluorescence sensor based on duplex-specific nuclease amplification for selective and sensitive detection of miRNAs in cancer cells
摘要： MicroRNAs (miRNAs) are considered as the potential biomarkers for many cancers. To determine miRNAs in cancer cells is significant for realizing these diseases. In this work, a microfluidic paper-based laser-induced fluorescence sensor based on duplex-specific nuclease (DSN) amplification was developed and applied to selectively and sensitively determine miRNAs in cancer cells. An interface for laser-induced fluorescence detection was firstly applied to perform the sample detection on the paper-based chip. Under the optimal conditions, DSN (3 μL 0.10 U) and Taqman probes (2 μL 2.5 × 10?7 M) were preserved on the circles (Diameter 4 mm) of the folded paper chip. When miRNA solution was added, the mixed solution could trigger fluorescence signal amplification by cyclically digesting hybrids of miRNAs and Taqman probes by DSN. The whole determination, including sample heating process, could be accomplished within 40 min. The detection limits for miRNA-21 and miRNA-31 were 0.20 and 0.50 fM respectively, corresponding to only 1.0 and 1.5 zmol consumption of miRNAs. The testing of mismatched miRNAs showed that the method had good specificity. Finally, the method was applied to determine miRNA-21 and miRNA-31 in lysates of cancer cells of A549 and HeLa, and hepatocyte LO2. MiRNA-21 and miRNA-31 could be successfully found from the two cancer cells. The concentrations for miRNA-21 and miRNA-31 were 1.74 × 10?13 M and 6.29 × 10?14 M in HeLa cell lysate (3.75 × 104 cells/mL), 3.07 × 10?15 M and 3.28 × 10?15 M in A549 cell lysate (8.33 × 106 cells/mL) respectively. The recoveries ranged from 87.30% to 111.83%, indicating the results were reliable. The developed method was effective, selective and sensitive in the determination of miRNAs in cancer cells.
关键词： microRNA,Duplex-specific nuclease,Paper-based sensor,Laser-induced fluorescence,Cancer cell
更新于2025-09-23 15:19:57
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Paper sensor of curcumin by fluorescence resonance energy transfer on nitrogen-doped carbon quantum dot
摘要： Paper Sensor detection methods are attractive in wide analytical applications. Presented herein is a paper sensor and ?uorescence methods that was ?rstly developed to detect curcumin (Cur) based on ?uorescence resonance energy transfer (FRET) between nitrogen-doped carbon quantum dots (NCQDs) and Cur. The facile ?uorescent method was demonstrated to detect Cur in the range of 0e2600 mM with a detection limit of 0.13 mM. And facile paper sensor of Cur was fabricated and displayed at concentration of 0 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM and 600 mM, respectively. In additions, it was realized for determination of Cur in real samples including orange juice and curry solution. Compared with the reported methods, the present method is simple, rapid and sensitive for detecting Cur.
关键词： Nitrogen-doped carbon quantum dots (NCQDs),Curcumin (Cur),Fluorescence resonance energy transfer (FRET),Paper-based sensor
更新于2025-09-11 14:15:04