Directional polymerisation fluorescent probe PCR and test kit

US12040049B2 · kind B2 · utility

Assignee

• Hong Jiang · CN

Inventors

• Hong Jiang · Nanhu, CN
• Yue Qu · Zhuhai, CN
• Chaoqi Qu · Zhuhai, CN

Key dates

• Filing date: Jul 2, 2019
• Grant date: Jul 16, 2024
• Expiry date: Jul 4, 2040

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y02A50/30
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A directional polymerisation fluorescent probe PCR and a test kit, wherein directional primer pair 5′ end complementary binding, sensitising, fluorescent probe 3′ end directional polymerisation and melting PCR are performed on the basis of primers and probe specific sequences, primer design uses pair side original primer 5′ end 5-10 bp reverse base sequences, added to primer pair front ends according to the 5′-3′ direction to form a “5′ reverse complementary sequence” chimeric primer pair for directional polymerisation, and 5′ complementarity enables amplification product 3′ ends to also be mutual templates and mutual primers for amplification. Sensitivity is increased by >2n geometric progression amplification, and competition decreases primer 3′ polymerisation PD non-specificity. Conventional hydrolysis fluorescent probe 3′ ends are added to probe central portion reverse complementary 5-8 bp end sequences to form two probe molecules for end portion-central portion close mutual hybridisation and directional polymerisation. Fluorescein-quenching groups at both ends of the probes are close to reduce reaction baseline fluorescence, and probe self-polymerisation inhibits primer polyme…