Biosensor for detecting repair glycosylase and detection method and application of biosensor

Abstract

The invention provides a biosensor for detecting repair glycosylase and a detection method and application of the biosensor. The biosensor comprises a stem ring DNA template and linear primer probes;the linear primer probes are LAMP primer probes, at least four primer probes are arranged, and comprises two front positive outer and inner primers, namely FIP and FOP, and two rear positive outer andinner primers BIP and BOP. A target base to be detected to repair glycosylase is designed in a stem region of the stem ring DNA template; and the stem ring DNA template is further provided with a region complementary to or identical to a primer probe sequence. The biosensor prepared by the invention actually constructs a self-directed replication system, and can be used for zero background, highsensitivity and high specificity detection of repair glycosylase activity. A simple, robust and universal platform is provided for repairing enzyme related biomedical research and early clinical diagnosis.

Description

technical field [0001] The invention belongs to the technical field of biological enzyme detection, and in particular relates to a biosensor for detecting and repairing glycosylase, a detection method and an application thereof. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Genomic DNA consists of Watson-Crick paired heterocyclic bases for the preservation and transmission of genetic information. Maintaining the accuracy and integrity of genomic DNA is therefore a fundamental prerequisite for all living organisms. However, genomic DNA is often damaged by various endogenous and exogenous factors, resulting in the production of about 10 5 damage (such as modified bases,...

AI Technical Summary

Problems solved by technology

PCR is a standard enzymatic DNA amplification technique based on thermocycling, but involves long assay times and precise thermocycling

Branched RCA and EXPAR are isothermal enzymatic amplification techniques, but require a variety of tool enzymes (ie, polymerase and nickase), and the operation is complicated and the experimental cost is high

In addition, PCR, branched RCA, and EXPAR are two or one primer-dependent amplification techniques, which cannot avoid cross-contamination caused by non-specific amplification

While LCR is based on thermostable DNA ligase-mediated adjacent repetitive cyclic ligation of hybridized DNA probes, the analysis of LCR products is always challenged by electrophoretic separation.

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