A DNA code-based multiplex SNV detection system, detection method and application

By combining LCR and HCR, specific recognition primers and hairpin probes were designed, and the products were enriched using magnetic microbeads. A DNA-encoded multiplex SNV detection system was constructed, which solved the problems of insufficient sensitivity, specificity and multiplex detection capability in the existing technology, and achieved efficient and low-cost SNV detection.

CN122279017APending Publication Date: 2026-06-26SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
Filing Date
2026-04-08
Publication Date
2026-06-26

AI Technical Summary

Technical Problem

Existing SNV detection technologies struggle to achieve an ideal balance between sensitivity, specificity, multiple detection capabilities, ease of operation, and cost-effectiveness, making it difficult to meet the demands for rapid, low-cost, or instant detection.

Method used

By combining ligase chain reaction (LCR) and hybridization chain reaction (HCR), and by designing specific recognition primers and hairpin probes, and using streptavidin-modified magnetic microbeads to enrich the products, a high-specificity recognition and signal amplification system based on DNA encoding was constructed.

Benefits of technology

It achieves high sensitivity and high specificity detection of multiple SNV targets in a single tube, can identify low-frequency variants in low-abundance samples, supports multiple parallel detection, and breaks through the limitations of existing technologies through multicolor fluorescent coding output, and can be applied to fields such as genetic disease screening and tumor mutation spectrum analysis.

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Abstract

This invention discloses a DNA-encoded multiplex SNV detection system, detection method, and application, belonging to the field of molecular diagnostics and gene detection technology. The system includes an SNV recognition and amplification module, a signal amplification and encoding module, and a product enrichment module. The detection method involves designing CP and TP probes targeting different SNV sites; CP and TP hybridize with target DNA in the sample; a thermostable DNA ligase catalyzes the formation of a ligation product between CP and TP under perfectly matched conditions; complementary probes HT1 and HT2 are introduced for thermal cycling amplification; fluorescently labeled hairpin probes H1 and H2 are added, triggering the HCR self-assembly to form a long-chain DNA polymer, outputting a detectable fluorescent signal. This invention achieves highly sensitive and specific detection of multiplex SNVs within a single tube through different fluorescent combinations, and is suitable for fields such as genetic disease screening and tumor mutation detection.
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