DNA insertion and deletion variation capture probes and uses thereof

By using heterologous hybridization double-stranded technology and specific capture probes combined with magnetic beads for separation, the problem of low-abundance DNA variants that are difficult to enrich by PCR technology has been solved, achieving efficient and sensitive variant detection.

CN116287121BActive Publication Date: 2026-06-19FUJIAN MATERNAL & CHILD HEALTH HOSPITAL
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
FUJIAN MATERNAL & CHILD HEALTH HOSPITAL
Filing Date
2023-03-29
Publication Date
2026-06-19

AI Technical Summary

Technical Problem

Existing PCR technology is difficult to efficiently and economically selectively enrich and detect low-abundance DNA insertion and deletion variants, and it is also difficult to simultaneously detect multiple DNA variant sites in the same reaction system.

Method used

The method employs heterologous hybridization double-stranded DNA to visualize insertion and deletion variations, utilizes specific capture probes to enrich heterologous hybridized double strands, and obtains variant DNA fragments through conventional PCR amplification. Finally, it combines biotin and minor groove binder-modified magnetic bead separation and purification techniques.

Benefits of technology

It enables simple, universal, efficient, and sensitive capture and detection of low-abundance DNA insertion and deletion variants, supporting high-throughput variant detection.

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Abstract

This invention provides a novel technique for capturing low-abundance DNA insertion and deletion variants. The capture probe sequence is constructed as 5'-CCGTGGAGAGACCACGG-(F)n-GGCACCAGAGAGGTGCC-3', comprising a 5' stem-loop region, a 3' stem-loop region, and a capture region (F)n between the stem-loop regions. The 5' stem-loop region sequence is 5'-CCGTGGAGAGACCACGG-3', and the 3' stem-loop region sequence is 5'-GGCACCAGAGAGGTGCC-3'. The capture region (F)n is a short tandem repeat fragment, F is the inverse complementary sequence of the inserted fragment or the deletion fragment sequence, and n is the number of short tandem repeats of the F fragment. The DNA insertion and deletion variant capture probe provided by this invention is simple, versatile, cost-effective, efficient, sensitive, and specific. It can effectively capture low-abundance insertion and deletion variant DNA fragments and perform high-throughput capture of unknown insertion and deletion variant DNA fragments, thereby enabling comprehensive and flexible detection of low-abundance insertion and deletion variant DNA fragments.
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