C-KIT gene multipoint mutation single tube fast detection method and kit

Abstract

The invention discloses a C-KIT gene multipoint mutation single tube fast detection method and kit. The C-KIT gene multipoint mutation single tube fast detection method comprises 1, designing ARMS primers and a corresponding downstream primer, intermediary connection primer, general fluorescent probe and quenching probe, 2, mixing the ARMS primers, the corresponding downstream primer, intermediary connection primer, general fluorescent probe and quenching probe and a hot-start fast Taq enzyme system and carrying out amplification, and 3, detecting a fluorescence change of the reaction system to determine if point mutation exists. The detection method can be operated simply and can realize qualitative detection of multipoint mutation existence. The detection method utilizes the primers with low design difficulty, greatly reduces a primer synthesis cost, solves limitation of the existing ARMS technology, greatly improves detection throughput, realizes single tube reaction replacing the existing multi-tube detection and saves manpower and material resources.

Description

technical field [0001] The invention relates to a mutation detection kit, in particular to a C-KIT multi-point mutation single-tube rapid detection kit. Background technique [0002] C-kit is a proto-oncogene, located on chromosome 4q11-12, with a size of about 80kb. The human C-kit receptor is a transmembrane receptor encoded by the C-kit proto-oncogene, with a total length of 976 amino acids and a relative molecular mass. The receptor has an extracellular ligand binding region, a transmembrane region, an intramembrane-adjacent region and a tyrosine protein kinase region (including TK1 and TK2), and belongs to the type III tyrosine kinase receptor family. [0003] Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the digestive tract, and it is also a model of tumor biological targeted therapy. Gastrointestinal stromal tumors originate from interstitial cells of Cajal (ICC) or primitive mesenchymal cells differentiated to ICC. [0004] The vast ...

AI Technical Summary

Problems solved by technology

[0006] The c-kit gene drug-sensitive mutation point is currently generally believed to need to detect 7 mutation points. The fluorescent PCR detection kits on the market generally use a single tube for detection, and the analysis process is cumbersome and unnecessary.

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