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Human epidermal growth factor receptor mutation gene detection kit

A technology of epidermal growth factor and detection kit, which is applied to the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problems of poor timeliness and high detection cost

Active Publication Date: 2014-10-08
安徽同科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, 8 PCR reactions need to be performed on the same sample at the same time to detect 29 mutant types, which is costly and time-sensitive

Method used

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  • Human epidermal growth factor receptor mutation gene detection kit
  • Human epidermal growth factor receptor mutation gene detection kit
  • Human epidermal growth factor receptor mutation gene detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Embodiment 1: the composition of kit

[0125] The composition of human epidermal growth factor receptor mutation gene (29 kinds) detection kit is shown in Table 2:

[0126] Table 2 Composition of detection kits for human epidermal growth factor receptor mutant genes (29 species)

[0127]

[0128]

[0129] Among them, the final concentration of each component in the PCR reaction solution A is 1×PCR buffer, 100nM each primer, 50nM each probe, dNTPs (0.4mM dATP, 0.4mM dGTP, 0.4mM dCTP, 0.2mM dTTP, 0.2mMdUTP) , 4mM MgCl 2 ;

[0130] In the present invention, the final concentration of each component in the PCR reaction solution A can also be 10×PCR buffer {(1mM or 10mM or 20mM or 50mM or 80mM or 100mM) KCl, (1mM or 10mM or 20mM or 50mM or 80mM or 100mM) (NH 4 ) 2 SO 4 , (1mM or 20mM or 50mM or 100mM or 150mM or 200mM) Tris-HCl (pH8 or pH8.5 or pH9)}, each primer (1nM or 10nM or 20nM or 50nM or 80nM or 90nM or 100nM), each probe ( 1nM or 10nM or 20nM or 50nM o...

Embodiment 2

[0136] Embodiment 2: the kit listed in embodiment 1 is used as human epidermal growth factor receptor mutant gene (29 kinds) detection Use of test kits

[0137] (1) DNA extraction

[0138] Take 30-50 mg of paraffin-embedded tissue samples. It is recommended to use TIANGEN Paraffin-embedded Tissue DNA Rapid Extraction Kit to extract genomic DNA. The extraction process is carried out according to the kit instructions, and finally 100 μL of DNA solution is collected.

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PUM

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Abstract

The invention relates to a human epidermal growth factor receptor mutation gene detection kit, which is characterized by comprising PCR (Polymerase Chain Reaction) reaction liquid A, PCR reaction liquid B, Taq / UNG enzyme mixed liquid, positive control and negative control, wherein the PCR reaction liquid A comprises 10*PCR buffer, a primer and a probe in deletion mutation of exon 19, a primer and a probe in T790M mutation, a primer and a probe in L861Q mutation, a primer and a probe of exon 2, MgC12, dNTPs and sterile distilled water; the PCR reaction liquid B comprises 10*PCR buffer, a primer and a probe in insertion mutation of exon 20, a primer and a probe of S768, a primer and a probe of L858R, a primer and a probe of G719X, MgC12, dNTPs and sterile distilled water. The human epidermal growth factor receptor mutation gene detection kit can be used for detecting human epidermal growth factor receptor mutation genes.

Description

technical field [0001] The invention belongs to the field of in vitro diagnostic reagents, in particular to a kit, and more specifically to a kit for detecting human epidermal growth factor receptor mutant genes (29 types). Background technique [0002] Non-small cell lung cancer (NSCLC) is a malignant tumor that seriously threatens human health. Despite the continuous improvement of surgery and chemotherapy techniques, the prognosis of patients is still poor, and the 5-year survival rate is less than 20%. At present, molecular targeted therapy targeting human epidermal growth factor receptor (EGFR) has become the most important way to treat NSCLC. [0003] EGFR is the expression product of the proto-oncogene C-erbB-1. The gene is located on chromosome 7 and belongs to the transmembrane receptor tyrosine kinase. After EGFR binds to its ligand, it can activate downstream signaling pathways, regulate tumor cell proliferation, differentiation, angiogenesis and apoptosis inhibi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113C12Q2563/107
Inventor 宋冬梅刘军辉刘代新
Owner 安徽同科生物科技有限公司