1. Detection of microRNAs by the combination of Exonuclease-III assisted target recycling amplification and repeated-fishing strategy
- Author
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Yao Chen, Zeng-Ping Chen, Xiao-Mei Yan, Ru-Qin Yu, and Yin-Qi Wang
- Subjects
Exonuclease III ,Detection limit ,biology ,Chemistry ,010401 analytical chemistry ,Biosensing Techniques ,DNA ,02 engineering and technology ,021001 nanoscience & nanotechnology ,01 natural sciences ,Biochemistry ,Fluorescence ,Gold foil ,0104 chemical sciences ,Analytical Chemistry ,MicroRNAs ,Exodeoxyribonucleases ,Limit of Detection ,biology.protein ,Environmental Chemistry ,Gold ,0210 nano-technology ,Biological system ,Nucleic Acid Amplification Techniques ,Spectroscopy - Abstract
A simple but effective method for the detection of miRNAs was proposed by integrating exonuclease-III assisted target recycling amplification and repeated-fishing strategy. In the proposed method, exonuclease-III assisted target recycling amplification reaction is adopted to produce a large amount of DNA fragments with fluorescence group at its 5′ end in the presence of the target miRNA, which are then repeatedly fished out from the reaction mixture by a gold foil modified with a capture probe and transferred into a so-called ‘product tube’. The amount of the target miRNA can then be determined from the fluorescence measurement of the solution in the ‘product tube’. Application to the detection of miRNA-155 in samples of KH-2 and BRSA-2B cells revealed that the proposed method could achieve sensitive and accurate quantification of the target miRNA with a limit of detection of 36 fM and recovery rates in the range from 96.2% to 105%. Its simplicity, sensitivity and resistance to possible fluorescence interferences in complex biological samples make the proposed method a potentially competitive alternative for miRNAs detection in complex biological samples.
- Published
- 2020