Method for detecting target mutation by carrying out retardation substitution amplification enrichment based on locked nucleic acid modified blocker

Abstract

The invention relates to a method for enriching and detecting gene mutation of a target region by utilizing locked nucleic acid modification. The enrichment method comprises the steps of mixing and amplifying a nucleic acid sample containing a to-be-detected target region, a first primer, a second primer and a retardation amplification sequence to obtain a first amplification product; and amplifying by utilizing the first primer and the third primer based on the first amplification product to obtain a second amplification product. The 3'terminal of the retardation amplification sequence contains a retardation group, wherein the retardation amplification sequence and the first primer contain partially overlapped sequences. The to-be-detected target region is located in a matching region ofthe retardation amplification sequence and the nucleic acid sample. The 3'terminal of the first primer is located at the upstream of the to-be-detected target region. The retardation amplification sequence is modified with locked nucleic acid, and a basic group carrying the locked nucleic acid is located at a position matching the to-be-detected target region. Therefore, high-precision detection of low-frequency gene mutation can be realized.

Description

technical field [0001] The present invention relates to the field of gene sequencing, in particular to a method for enriching and detecting gene mutations in a target region by using locked nucleic acid modification, and in particular to a method for blocking, replacing, amplifying, enriching and detecting targets based on locked nucleic acid modified blockers method of mutation. Background technique [0002] With the rapid development of the times and technology, the concept of cancer treatment is undergoing fundamental changes. The International Union Against Cancer believes that 1 / 3 of cancers can be prevented, 1 / 3 of cancers can be cured if diagnosed early, and 1 / 3 of cancers can be treated correctly to relieve pain and prolong life. The treatment of cancer has also changed from empirical science to evidence-based medicine, from cell attack mode to targeted treatment mode. Early traditional treatment methods were surgical resection, chemotherapy and radiotherapy, among...

AI Technical Summary

Problems solved by technology

Therefore, there is an urgent need for a ctDNA detection technology that can not only detect low-frequency mutation sites but also achieve a certain throughput. None of the main ctDNA detection technologies on the market can meet the requirements.

This kind of low-frequency mutation site detection with a certain throughput is a bottleneck encountered by current ctDNA detection technology

Images