Primer and method for detecting human EGFR gene mutation

A technology of human and external primers, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems of electrophoretic bands that cannot be displayed, low selectivity, and low amplification efficiency. Achieve the effect of fast detection speed, strong selectivity and high sensitivity

Active Publication Date: 2014-11-05
艾吉泰康(嘉兴)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ARMS method can increase the selectivity of mutation detection to 1%, but for DNA samples with low quality or low content, the selectivity is still low
In addition, the ARMS method relies on conventional gel electrophoresis or probe hybridization to detect PCR products. Mutation products with low amplification efficiency due to low sample content may not be displayed on the electrophoresis band.
At the same time, this method cannot perform quantitative or semi-quantitative analysis of the mutations contained in the sample

Method used

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  • Primer and method for detecting human EGFR gene mutation
  • Primer and method for detecting human EGFR gene mutation
  • Primer and method for detecting human EGFR gene mutation

Examples

Experimental program
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Effect test

Embodiment 1

[0090] Detection of single mutations in paraffin-embedded histopathological sections of non-small cell lung cancer. In this example, a series of primers were designed using the technology of the present invention, and combined with capillary electrophoresis to detect the gene mutation of the EGFR gene in the sample, that is, the 858th mutation site of exon 21 of the Epidermal Growth Factor Receptor (Epidermal Growth Factor Receptor).

[0091] The experimental materials are as follows:

[0092] Reagent manufacturers rTaq enzyme TAKARA (treasure biological) company 10× buffer TAKARA (treasure biological) company dNTP-MIX TAKARA (treasure biological) company MgCl 2 TAKARA (treasure biological) company Primer synthesis Beijing Qingke Xinye Biotechnology Co., Ltd. Formamide American SIGMA Corporation DMSO American SIGMA Corporation Proteinase K QIAGEN Corporation QIAamp MinElute column QIAGEN Corporation ...

Embodiment 2

[0133] Detection of deletions in paraffin-embedded histopathological sections of non-small cell lung cancer. In this embodiment, a series of primers are designed with the technology of the present invention, and combined with capillary electrophoresis to detect the EGFR gene in the sample, that is, the gene at the point E746 (2235-2249 DEL15 ) of exon 19 of the epidermal growth factor receptor (Epidermal Growth Factor Receptor) missing.

[0134] The experimental materials are as follows:

[0135] Reagent manufacturers rTaq enzyme TAKARA (treasure biological) company 10× buffer TAKARA (treasure biological) company dNTP-MIX TAKARA (treasure biological) company MgCl 2 TAKARA (treasure biological) company Primer synthesis Beijing Qingke Xinye Biotechnology Co., Ltd. Formamide American SIGMA Corporation DMSO American SIGMA Corporation Proteinase K QIAGEN Corporation QIAamp MinElute column QIAGEN Corporation...

Embodiment 3

[0176] Detection of single mutations in paraffin-embedded histopathological sections of non-small cell lung cancer. In this example, a series of primers were designed using the technology of the present invention, and combined with capillary electrophoresis to detect the gene mutation of the EGFR gene in the sample, that is, the T790M mutation site of exon 20 of the Epidermal Growth Factor Receptor (Epidermal Growth Factor Receptor).

[0177] The experimental materials are as follows:

[0178] Reagent manufacturers rTaq enzyme TAKARA (treasure biological) company 10× buffer TAKARA (treasure biological) company dNTP-MIX TAKARA (treasure biological) company MgCl 2 TAKARA (treasure biological) company Primer synthesis Beijing Qingke Xinye Biotechnology Co., Ltd. Formamide American SIGMA Corporation DMSO American SIGMA Corporation Proteinase K QIAGEN Corporation QIAamp MinElute column QIAGEN Corporation ...

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Abstract

The invention belongs to the field of biotechnology, especially the detection of gene mutation. More specifically, the invention discloses 9 groups of primers for detection of 41 mutation of a human epidermal growth factor EGFR gene and a detection method by using the primers.

Description

[0001] technical field [0002] The invention relates to the detection of gene mutation, in particular to a primer and a method for detecting the mutation of epidermal growth factor EGFR gene. Background technique [0003] Epidermal Growth Factor Receptor (EGFR) is a receptor for epidermal growth factor (EGF) cell proliferation and signal transduction. EGFR is a member of the HER or ErbB receptor family, which includes EGFR (ErbB-1), HER2 / c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR is also often referred to as HER1 or ErbB1. EGFR and its family members can regulate many physiological processes such as cell differentiation, apoptosis, cell movement, angiogenesis, invasion and metastasis, and are expressed in most human tumors. Therefore, EGFR has been proved to be effective in many tumors. It plays an important regulatory role in the process of occurrence, development and prognosis. [0004] EGFR is a typical receptor with Tyrosine Kinase (TK) activity, and TK ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6858C12Q2535/137C12Q2565/125
Inventor 不公告发明人
Owner 艾吉泰康(嘉兴)生物科技有限公司
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