Fluorescence chemical sensor and method for simultaneously detecting diversified DNA (deoxyribonucleic acid) glycosylases on single-molecular levels and application of fluorescence chemical sensor

Abstract

The invention discloses a fluorescence chemical sensor and a method for simultaneously detecting diversified DNA (deoxyribonucleic acid) glycosylases on single-molecular levels and application of thefluorescence chemical sensor. The fluorescence chemical sensor, the method and the application have the advantages that the fluorescence chemical sensor is based on two different molecular beacons including a molecular beacon modified by 8-hydroxyl guanine and a molecular beacon modified by deoxygenated hypoxanthine, the tail end of the molecular beacon modified by the 8-hydroxyl guanine is labeled by cyanine 3 (Cy3) and quenching groups, the tail end of the molecular beacon modified by the deoxygenated hypoxanthine is labeled by cyanine 5 (Cy5) and quenching groups, the fluorescence chemicalsensor is used for detecting 8-hydroxyl guanine DNA glycosylases and N-methylpurine DNA glycosylases and is different from the traditional molecular beacons which can be severely affected by dynamicsand thermodynamics, signal restoration of the Cy3 and the Cy5 depends on molecular beacon splitting with the DNA glycosylases used as media, the DNA glycosylases can be simultaneously sensitively detected by the aid of the method without optional signal amplification, the activity of hOGG1 and hAAG can be detected by the aid of the method in an ultra-sensitive manner without optional signal amplification, the fluorescence chemical sensor can be easily, conveniently and quickly operated, and accurate and reliable test results can be obtained.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a fluorescent chemical sensor based on a single-molecule level for simultaneously detecting multiple DNA glycosylases, a detection method and an application thereof. Background technique [0002] DNA glycosylase is an initial enzyme in the base excision repair pathway. This enzyme can recognize and excise mutated DNA bases and generate apurine or apyrimidine (AP) sites. The normal DNA glycosylase The activity ensures the stability of base excision repair and thus the stability of the genome. The base excision repair pathway involves at least 11 different mammalian glycosylases that operate by relying on different mutation types. In addition, abnormal DNA glycosylase is closely related to a series of diseases such as cancer, neuropathy, cardiovascular disease and inflammation. [0003] Traditional methods for detection of glycosylases include radiolabelin...

AI Technical Summary

Problems solved by technology

These methods improve the detection sensitivity through signal amplification through amplification, but exonuclease-assisted signal amplification requires high reaction system and poor reproducibility

However, target-mediated autocatalytic DNAzyme-generated rolling circle amplification is prone to non-specific amplification, resulting in false positive signals and reduced specificity.

At the same time, low denaturation temperature polymerase chain reaction relies on high-precision temperature cycling, a variety of primers and specific DNA polymerases, increasing the complexity and cost of detection

In addition, the above methods of amplifying the signal can only be used to measure a DNA glycosylase

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