Method for simultaneously detecting multiple miRNAs through fluorescence method

Abstract

The invention discloses a method for simultaneously detecting the concentration of multiple miRNAs through a fluorescence method. The method comprises the steps of selecting miRNAs; preparing a fluorescein-labelled nucleic acid probe; selecting a fluorescence donor; calculating fluorescent crosstalk correction factors of fluorescein corresponding to the multiple miRNAs; obtaining a corresponding relationship between the fluorescence intensity of the corrected fluorescein corresponding to the multiple miRNAs and the concentration of the miRNAs; and detecting fluorescence signals of the fluorescein corresponding to the multiple miRNAs in a to-be-detected miRNA solution and calculating the concentration of the miRNAs by using the corresponding relationship between the fluorescence intensity of the corrected fluorescein corresponding to the multiple miRNAs and the concentration of the miRNAs. Simultaneous detection of multiple miRNAs is achieved by using single wavelength excitation; the detection method is simplified; the interference between a background signal and fluorescein signals is effectively reduced; the concentration of any miRNA in the solution can be accurately measured; and the method is low in cost, fast, simple, sensitive and good in specificity.

Description

technical field [0001] The invention belongs to the technical field of biosensing, and in particular relates to a method for simultaneously detecting multiple miRNAs through a fluorescence method. Background technique [0002] Lung cancer is the leading cause of cancer death worldwide, with more than 1 million deaths due to lung cancer each year, of which non-small cell lung cancer (NSCLC) accounts for more than 80%, including adenocarcinoma, squamous cell carcinoma and large cell carcinoma. cancer. Although great progress has been made in lung cancer imaging examination and targeted therapy in recent years, the 5-year overall survival rate of NSCLC is still less than 15%. The lack of effective early diagnosis is the main reason for the poor prognosis of NSCLC. Most patients are already in the middle and advanced stage when they are clinically diagnosed, and have lost the opportunity for radical surgery. Examinations such as chest X-ray and CT have low specificity, while ...

AI Technical Summary

Problems solved by technology

Although great progress has been made in lung cancer imaging examination and targeted therapy in recent years, the 5-year overall survival rate of NSCLC is still less than 15%.

However, the analytical detection of miRNAs remains challenging due to their unique properties, including short size, high sequence similarity (homology), tolerance to degradation, etc., making miRNAs more difficult to apply to nucleic acid hybridization-based biological Therefore, in the field of sensors, there is an urgent need to develop a series of methods for rapid, sensitive and multi-channel detection of miRNA in the future.

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