Primer, detection method and kit for detecting human PIK3CA gene E545K mutation

Abstract

The invention provides a primer, detection method and kit for detecting human PIK3CA gene E545K mutation. The detection primer provided by the invention can be combined with a specific sequence of a PIK3CA gene E545K to amplify a mutation sequence. The detection method provided by the invention can be combined with an amplified fragment by adopting a fluorescence signal detection group to send outa fluorescence signal; a blocking agent is specifically combined with a wild type sequence corresponding to a mutation site to inhibit wild type non-specific amplification. A competitive allele specific PCR (Polymerase Chain Reaction) amplification technology is adopted to establish the kit based on real-time fluorescence PCR and the PIK3CA gene E545K mutation can be accurately detected. The method provided by the invention has the advantages of simplicity in operation, easiness of reading results and high sensitivity, and can be used for detecting a sample containing 0.001 percent of the PIK3CA gene E545K mutation; ultrasensitive detection of the PIK3CA gene E545K mutation is realized.

Description

[0001] This application claims the priority of the Chinese patent application submitted to the China Patent Office on September 08, 2017, with the application number 201710805686.5, and the title of the invention is "Primers, detection methods and kits for ultrasensitive detection of human PIK3CA gene E545K mutation" rights, the entire contents of which are incorporated in this application by reference. technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a primer, a detection method and a kit for detecting the E545K mutation of human PIK3CA gene. Background technique [0003] Point mutations are closely related to many tumors. Studies have shown that point mutations at certain gene sites can greatly increase the incidence of tumors. Phosphatidylinositol-3 kinase (PI3K) is a kinase that catalyzes the phosphorylation of inositol groups. It is a heterodimer composed of a catalytic subunit and a regulatory subunit; it is div...

AI Technical Summary

Problems solved by technology

[0005] At present, gene mutation detection can start from two levels: one is the indirect detection at the protein level, which mainly uses some protein analysis techniques to analyze the mutant protein produced by the mutant and other changes caused by the mutant protein; the other is the genetic material (mainly It is the direct detection of deoxyribose nucleic acid (DNA), mainly represented by polymerase chain reaction (Polymerase chain reaction, PCR) technology, the detection accuracy of this technology is consistent with the results of tissue samples; the sequencing method can directly obtain the target The base sequence of a DNA fragment, but it is time-consuming and laborious compared to many mutation detection methods

Images