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Sequence composition and kit for detecting mutation of human EGFR gene

A technology of composition and kit, which is applied in the field of sequence composition and kit for detecting human EGFR gene mutation, which can solve the problems of low sensitivity and detection throughput, limited PCR detection channel, and difficulty in simultaneous detection of EGFR mutation gene, etc. problems, to achieve the effect of avoiding false positive signals, strong specificity, and high sensitivity

Active Publication Date: 2018-12-07
SHANGHAI MAG GENE NANOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the ARMS-PCR method is a relatively simple and highly sensitive detection method, it is difficult to simultaneously detect multiple EGFR mutation genes due to the limited PCR detection channel.
[0008] Therefore, in order to simultaneously solve the problems of low sensitivity and detection throughput faced by the existing EGFR mutation gene detection technology, the present invention is devoted to developing a kind of EGFR gene mutation The primer set, blocker set and their use in modified-ARMsPCR

Method used

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  • Sequence composition and kit for detecting mutation of human EGFR gene
  • Sequence composition and kit for detecting mutation of human EGFR gene
  • Sequence composition and kit for detecting mutation of human EGFR gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Detection of E746-A750del on exon 19 of EGFR gene hotspot mutation

[0038] This method mainly includes the following steps:

[0039] (1) Synthesize wild-type gene sequences and corresponding mutant gene sequences based on exons 18, 19, 20, and 21 of EGFR, and design corresponding primers and blockers. The detailed sequences are shown in Table 3. Among them, SEQ ID1-SEQ ID3 can simultaneously amplify the mutation type comprising 19DEL; SEQ ID4-SEQ ID8 can simultaneously amplify the mutation type comprising G719A / G719S / G719C; SEQ ID9-SEQ ID15 can simultaneously amplify the mutation type comprising 20insert ; SEQ ID16-SEQ ID18 can simultaneously amplify the mutation type comprising L858R; SEQ ID19-SEQ ID21 can simultaneously amplify the mutation type comprising T790M; SEQ ID22-SEQ ID24 can simultaneously amplify the mutation type comprising S768I; SEQ ID25-SEQ ID27 can simultaneously amplify the mutation type containing L861Q;

[0040] (2) Extract template DN...

Embodiment 2

[0044] The detection of the S768I / L861Q point mutation of embodiment 2.EGFR gene

[0045] This method mainly includes the following steps:

[0046] (1) Synthesize wild-type gene sequences and corresponding mutant gene sequences based on exons 18, 19, 20, and 21 of EGFR, and design corresponding primers and blockers. The detailed sequences are shown in Table 3. Among them, SEQ ID1-SEQ ID3 can simultaneously amplify the mutation type comprising 19DEL; SEQ ID4-SEQ ID8 can simultaneously amplify the mutation type comprising G719A / G719S / G719C; SEQ ID9-SEQ ID15 can simultaneously amplify the mutation type comprising 20insert ; SEQ ID16-SEQ ID18 can simultaneously amplify the mutation type comprising L858R; SEQ ID19-SEQ ID21 can simultaneously amplify the mutation type comprising T790M; SEQ ID22-SEQ ID24 can simultaneously amplify the mutation type comprising S768I; SEQ ID25-SEQ ID27 was able to simultaneously amplify mutation types containing L861Q;

[0047] (2) Extract template D...

Embodiment 3

[0051] Example 3. Detection of insertion mutation (ID39) on exon 20 of EGFR gene hotspot mutation

[0052] This method mainly includes the following steps:

[0053] (1) Synthesize wild-type gene sequences and corresponding mutant gene sequences based on exons 18, 19, 20, and 21 of EGFR, and design corresponding primers and blockers. The detailed sequences are shown in Table 3. Among them, SEQ ID1-SEQ ID3 can simultaneously amplify the mutation type comprising 19DEL; SEQ ID4-SEQ ID8 can simultaneously amplify the mutation type comprising G719A / G719S / G719C; SEQ ID9-SEQ ID15 can simultaneously amplify the mutation type comprising 20insert ; SEQ ID16-SEQ ID18 can simultaneously amplify the mutation type comprising L858R; SEQ ID19-SEQ ID21 can simultaneously amplify the mutation type comprising T790M; SEQ ID22-SEQ ID24 can simultaneously amplify the mutation type comprising S768I; SEQ ID25-SEQ ID27 was able to simultaneously amplify mutation types containing L861Q;

[0054] (2) E...

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Abstract

The invention discloses a group of primer sequences and blocker sequences for detecting the mutation of a human EGFR gene and relates to detection of the mutation of the human EGFR gene. The inventionfurther discloses a kit including the primer sequences and blocker sequences and a use method of the kit. The primer sequences and blocker sequences provided by the invention can be used for detecting multiple different types of mutant genes in a single reaction and are high in sensitivity, strong in specificity and high in single detection flux.

Description

technical field [0001] The invention relates to a method for detecting human gene mutations, in particular to a sequence composition for detecting human EGFR gene mutations, a kit and a method thereof. Background technique [0002] Human epidermal growth factor (epidermal growth factor receptor, EGFR) is a protein tyrosine kinase receptor, which is widely distributed on the surface of mammalian epithelial cells, fibroblasts, glial cells and other cells, and has tyrosine kinase activity. EGFR is one of the members of HER / ErbB family. After EGFR binds to its ligand, it forms a dimer on the cell surface, activates receptor autophosphorylation through the activity of tyrosine kinase, and regulates transcription factors to activate the transcription of genes through the cascade reaction of adapter proteins and enzymes in the cytoplasm. Directs cell migration, proliferation, differentiation and apoptosis. When EGFR is mutated, EGFR itself or its ligands are overexpressed, thereb...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2537/143C12Q2531/113C12Q2535/137C12Q2537/163
Inventor 徐高连徐宏古宏晨
Owner SHANGHAI MAG GENE NANOTECH CO LTD
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