Rapid single-tube detection method and rapid single-tube detection kit for multipoint mutation of PDGFRA (platelet-derived growth factor receptor alpha) gene

Abstract

The invention discloses a rapid single-tube detection method and a rapid single-tube detection kit for multipoint mutation. The rapid single-tube detection method for multipoint mutation includes: 1) designing ARMS primers as well as downstream primers, mediated ligation primers, a universal fluorescence probe and a quenching probe corresponding to the ARMS primers; 2) mixing the ARMS primers as well as the downstream primers, the mediated ligation primers, the universal fluorescence probe and the quenching probe corresponding to the ARMS primers with a rapid hot-start Taq enzyme system prior to amplification; 3) detecting fluorescence change of a reaction system and determining whether point mutation exists or not. The rapid single-tube detection method has the advantages that the rapid single-tube detection method is simple to operate and capable of detecting existence of multipoint mutation qualitatively; the primers used in the detection method are low in design difficulty, so that primer synthesis cost is reduced substantially; limitations of an existing ARMS technology are overcome, and detection throughput is improved greatly; substituting single-tube reaction for previous multi-tube detection is achieved, and accordingly, manpower and material resources are saved.

Description

technical field [0001] The invention relates to a mutation detection kit, in particular to a PDGFRA multi-point mutation single-tube rapid detection kit. Background technique [0002] Platelet-derived growth factor receptor (PDGFR) is an important member of the PDGFR signal transduction pathway, including PDGFRalpha and PDGFRbeta, which play an important role in promoting tumor cell proliferation, invasion, and neovascularization. ,PDGFRA gene mutation is mainly seen in gastrointestinal stromal tumor (GIST), so it is often used as a reference index for the diagnosis of GIST. [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 tyrosine kinase inhibitor imatinib mesylate (Gleevec) can selectively inhibit the kinase...

AI Technical Summary

Problems solved by technology

[0005] The mutation points sensitive to PDGFRA gene drugs are currently generally believed to need to detect 4 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.

Images