Rapid detection method for nanometer fluorescent trace of specific nucleic acid fragment on basis of CRISPR-Cas12g

Abstract

The invention discloses a rapid detection method for nanometer fluorescent trace of specific nucleic acid fragment on the basis of CRISPR-Cas12g. The rapid detection method comprises the following steps: with the specific nucleic acid fragment in a biological sample as a research object, preparing a nucleic acid target, a quenching fluorescence probe and a Cas12g ternary complex of the biologicalsample; mixing the Cas12g ternary complex, the nucleic acid target, the quenching fluorescent probe, background RNA and an RNA enzyme inhibitor in a nuclease buffer solution for incubation, after theCas12g ternary complex is recognized and matched with the nucleic acid target, shearing the nucleic acid target in accompany with non-specific trans-shearing of the quenching fluorescent probe so as to obtain detectable fluorescence; and collecting fluorescence spectrum data, and constructing a quantitative detection model of an up-conversion nanometer fluorescence intensity change value and nucleic acid targets with different concentrations so as to realize rapid detection of the nanometer fluorescence trace of the nucleic acid target in the biological sample. The rapid detection method provided by the invention is applicable to rapid, specific and sensitive detection of the specific nucleic acid fragment in the biological sample.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, in particular to a rapid detection method for nano-fluorescent traces of specific nucleic acid fragments based on CRISPR-Cas12g. Background technique [0002] With the rapid development of nucleic acid detection technology, rapid detection methods for various biological samples have emerged one after another. For example, food-borne pathogenic bacteria that cause food-borne diseases have become a hot issue in food safety in the world. In recent years, the threat of bioterrorism and foodborne disease incidents at home and abroad have aroused people's high attention to foodborne pathogenic bacteria. Rapid detection of specific nucleic acid fragments of foodborne pathogens in biological samples is an important means to prevent foodborne diseases. [0003] At present, nucleic acid detection technologies include PCR-based detection technologies (common PCR, multiplex PCR, real-time fl...

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Problems solved by technology

At home and abroad, there are no reports on the use of CRISPR-Cas12g system to achieve nano-fluorescence quantitative detection of specific nucleic acid fragments in biological samples

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