Journal
TALANTA
Volume 146, Issue -, Pages 648-654Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2015.06.060
Keywords
MicroRNA; Lateral flow biosensor; Visual detection; Signal amplification; Horseradish peroxidase
Categories
Funding
- National Natural Science Foundation of China (NSFC) [21475009, 21475008, 21127007, 21275017]
- Fundamental Research funds for the Central Universities [FRF-TP-14-065A2, FRF-TP-14-015A1]
- Beijing Higher Education Young Elite Teacher Project [YETPO424]
- China Postdoctoral Science Foundation [2014M550610]
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An enzyme-based dual-labeled nanoprobe is designed to fabricate a sensitive enzyme-amplified lateral flow biosensor for visual detection of mircoRNA-224 (miRNA-224). The recognition DNA probe (detection probe) and signal amplification enzyme (Horseradish peroxidase, HRP) are immobilized on gold nano-particle (GNPs) surface, simultaneously. The capture DNA probes are immobilized on the test zone of the lateral flow biosensor. When miRNA-224 is present, the enzyme-based dual-labeled nanoprobes will be captured by forming the sandwich structure on the test zone of the lateral flow biosensor, enabling the visual detection for miRNA-224. Sensitivity is amplified by applying the 3,3,5,5-tetramethylbenzidine enzymatic substrate (TMB/H2O2 enzymatic substrate) onto the test zone. The enzymatic reactions between the HRP and the TMB/H2O2 enzymatic substrate will produce blue products, which deposit on the nanoprobe surface to enhance the visual effect and the corresponding response intensities of the test zone. This enzyme-amplified lateral flow biosensor shows a low limit of detection (LOD) (7.5 pM) toward miRNA-224 in the buffer solution, which is improved by 10-fold than that of the single-labeled lateral flow biosensor. This biosensor has been successfully used for the detection of the target miRNA-224 detection in A549 cell lysate. (C) 2015 Elsevier B.V. All rights reserved.
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