Primer for detecting human gastrointestinal stromal tumor C-KIT gene V559A mutation, detection method and kit thereof

Abstract

The invention provides a primer for detecting human gastrointestinal stromal tumor C-KIT gene V559A mutation, a detection method and a kit thereof. The detection primer provided by the invention can bind to a specific sequence of the C-KIT gene, and amplifies a mutant sequence. The detection method adopts a detection fluorescent signal group to bind to an amplified fragment to emit a fluorescent signal, and uses a blocking agent specifically binding to the wild type sequence corresponding to the mutation site to inhibit wild type non-specific amplification. The method adopts a competitive allele specific PCR amplification technology to establish a real-time fluorescent PCR kit, which can accurately detect the C-KIT gene V559A mutation. The method of the invention has simple operation, easyresult reading, and high sensitivity, can detect a sample containing 0.001% C-KIT gene V559A mutation, and realizes ultrasensitive detection of the C-KIT gene V559A mutation.

Description

[0001] This application requires the application number submitted to the Chinese Patent Office on September 08, 2017. The application number is 201710806478.7, and the title of the invention is "Primers, detection methods and kits for ultrasensitive detection of C-KIT gene mutations in human gastrointestinal stromal tumors." The priority of the Chinese patent application ", the entire content of which is incorporated in this application by reference. technical field [0002] The invention belongs to the field of biological technology, and in particular relates to a primer, a detection method and a kit for detecting the V559A mutation of the human gastrointestinal stromal tumor C-KIT gene. Background technique [0003] C-KIT is a proto-oncogene, located on chromosome 4q11-12, with a size of about 80kb, encoding a 145KD transmembrane protein CD117, belonging to the class III tyrosine kinase receptor family, with an extracellular ligand binding region, transmembrane region, int...

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Problems solved by technology

However, due to the low content of oncogenes in early cancer samples, especially blood samples, the detection sensitivity of the above methods can only reach 0.2%, which is far from the detection of extremely small amounts of abnormal DNA molecules from a large amount of normal DNA background. Require

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