Quantitative detection method for specific nucleic acid fragments based on CRISPR technology

Abstract

The invention discloses a quantitative detection method for specific nucleic acid fragments based on the CRISPR technology. The quantitative detection method comprises the following steps: acquiring atreated target nucleic acid sample, and adding the target nucleic acid sample and an aqueous-phase solution into a droplet generation device to generate reaction droplets, wherein the reaction droplets comprise at least one nucleic acid fragment to be tested; amplifying the nucleic acid fragment to be tested in the reaction droplets by using a recombinase polymerase amplification technique, and allowing target gene signals in the nucleic acid fragment to be tested to realize cascade amplification by using a gene editing technique; collecting a positive signal corresponding to a target gene, and converting the positive signal into image data; processing a fluorescence threshold in the image data by using the Poisson distribution principle so as to obtain the test corresponding to the number of the target gene, corresponding to the positive signal, in the nucleic acid fragment to be tested. Thus, rapid efficient quantitative detection of the concentration of the target gene is realizedbased on the CRISPR technology.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method for quantitative detection of specific nucleic acid fragments based on CRISPR technology. The method for quantitative detection of specific nucleic acid fragments can be applied to quantitatively detect specific nucleic acid fragments quickly and efficiently. Background technique [0002] With the development of science and technology and the improvement of medical level, people have made practical progress in the research on the correlation between various diseases and gene mutations or polymorphisms, and gradually understand the relationship between gene mutations or polymorphisms and single gene diseases , the relationship between multifactorial diseases. Based on the relationship between genes and diseases, people can easily and accurately detect diseases by detecting gene mutations in the target gene or the region containing the target gene, and the detection of...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a quantitative detection method for specific nucleic acid fragments based on CRISPR technology. The quantitative detection method for specific nucleic acid fragments solves the problem of quantitative detection of nucleic acid fragments, and can quickly and efficiently detect single-molecule specific nucleic acid fragments quantitatively.

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