A method for detecting esr1 gene mutation based on fluorescent PCR technology

Abstract

The invention provides a method for rapid and accurate detection of four mutations Y537S, Y537C, Y537N and D538G of an ESR1 gene based on fluorescent PCR, wherein the method can avoid false result generation, is high in sensitivity and wide in adaptation range, and saves consumable materials and time. Upstream primers respectively designed aiming at the four mutations comprise 5'-GAACGTGGTGCCCCTCcC-3', 5'-GAACGTGGTGCCCCTgTG-3', 5'-AGAACGTGGTGCCCCaCA-3', and 5'-ACGTGGTGCCCCTCTATtG-3'; a designed common downstream primer is 5'-CACGGCTAGTGGGCGC-3; and a designed common TaqMan fluorescence probe is 5'-FAM-CTGGAGATGCTGGACGCCCAC-BHQ2-3'.

Description

technical field [0001] The invention relates to a method for detecting ESR1 gene mutation. Background technique [0002] Human estrogen receptor α (Estrogenreceptoralpha, ESRα) is encoded by the ESR1 (EstrogenReceptor1) gene. After combining with its ligand estrogen, ESRα can activate a series of cell biological effects in cells. The mutation and abnormal expression of ESR1 gene are related to the occurrence of breast cancer and other diseases. [0003] The most common mutation types of ESR1 gene are Y537S, Y537C, Y537N and D538G. These mutations alter the conformation of ESRα, which can be constitutively activated in the absence of estrogen binding. [0004] The currently reported methods for detecting ESR1 mutations are direct sequencing technology and next-generation sequencing technology. Among them, the direct sequencing technology is low in cost and high in accuracy. However, it requires PCR post-processing steps, which is complicated to operate and prone to contam...

AI Technical Summary

Problems solved by technology

Among them, the direct sequencing technology is low in cost and high in accuracy, but requires PCR post-processing steps, which is not only complicated to operate, but also prone to contamination and false results; and this method is not sensitive enough to fully adapt to the complex situation of clinical samples

The next-generation sequencing technology has high sensitivity, but it relies on expensive instruments and analysis software, which greatly increases the detection cost and is difficult to be used as a universal technology

Its core principle is that the primer extension guided by DNA polymerase starts from the 3' end of the primer in the PCR process, and the degree of complementary pairing between the base at the 3' end of the primer and the template strongly affects the recognition of the polymerase and the progress of the PCR reaction: If the base is normally complementary to the template (A-T, G-C), the primer can be extended without interruption, and PCR can be efficiently performed to obtain a complete product; otherwise, if the base is not normally paired with the template, the extension of the primer is hindered stagnation, PCR efficiency is greatly reduced

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