Objective:To established a high sensitivity method based on nucleic acid probe hybridization and enzyme-linked chromogenic for qualitative and quantitative detection of microRNA-21(miRNA-21). Methods:Biotin labelled miRNA-21 capture probe was immobilized on 96-well plate by biotin-streptavidin system. The capture probe could effectively bind to the target miRNA-21 which appeared in the reaction system. DNA detection probe labeled by horseradish peroxidase which had catalytic activity and signal amplification could catalytic substrate 3, 3', 5, 5'-tetramethylbenzidine to form a colorimetric reaction. Qualitative and quantitative analysis of the miRNA-21 concentration were performed based on the color change of reaction system. Result:By investigating the sensitivity,precision and recovery of the method, the lowest detection limit was 0.81 pmol·L-1, relative standard deviationwere less than 7.5%, and the recoveries were between 78.9%-119.2%.Conclusion:In this study, a sensitive and high throughput chromogenic method for detection of miRNA-21 is established. It has good sensitivity, repeatability and recovery. |
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