Primers capable of simultaneously acquiring and detecting multiple target area sequences, kit and method

Abstract

The invention provides multiple PCR primers capable of amplifying and / or detecting target gene areas, such as amplifying and detecting mutant sites of EGFR, KRAS and / or BRAF genes. By using the multiple PCR primers, the mutant sites of EGFR / KRAS / BRAF genes can be efficiently detected in one time, and a plurality of exons on the EGFR, KRAS and / or BRAF genes can be simultaneously covered.

Description

[0001] related application [0002] This application is a divisional case of Chinese patent application 201510740906.1 with an application date of November 4, 2015 and titled "Multiple PCR primers and methods and applications for detecting EGFR / KRAS / BRAF gene mutation sites based on next-generation sequencing technology" . technical field [0003] The invention belongs to the field of molecular biology, in particular to a multiplex PCR primer, method and application for detecting EGFR, KRAS and / or BRAF gene mutation sites based on next-generation sequencing technology. Background technique [0004] Epidermal growth factor receptor (EGFR) is the expression product of the proto-oncogene C-erbB-1 (HER-1) located on the short arm of chromosome 7, and is a member of the epidermal growth factor receptor family (HER). One of the four members of the HER family, the HER family plays an important regulatory role in cellular physiological processes. Mutations in the EGFR tyrosine kin...

AI Technical Summary

Problems solved by technology

In the Sanger sequencing method, a single pair of primers can detect multiple mutations, but the mutation sites on multiple genes or different exons of the same gene need to be amplified and sequenced separately, which is cumbersome and has a low sensitivity of about 20 %, higher false negative rate

Although the fluorescent quantitative PCR method has high sensitivity, each pair of primers can only detect one mutation, and each mutation requires a separate PCR reaction system, and the simultaneous detection of multiple mutation sites in multiple samples is cumbersome.

These two methods require a large amount of samples and are not suitable for detecting multiple gene mutation sites at the same time

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