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EGFR genotyping detection method based on nucleic acid tetrahedron probe modified printed gold electrode and kit

A tetrahedral probe and detection kit technology, used in biochemical equipment and methods, microbial determination/inspection, material analysis by electromagnetic means, etc., can solve the problems of insufficient sensitivity and low specificity, and improve the sensitivity , high sensitivity, less sample loading

Inactive Publication Date: 2021-09-10
王旭耀
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Problems solved by technology

[0005] The problem that the present invention solves is to provide a kind of detection method and kit for two main mutation types (19del and L858R) in the EGFR gene mutation based on nucleic acid tetrahedral structure probe modification printing gold electrode, to overcome the prior art sensitivity Insufficient, low specificity and other shortcomings

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  • EGFR genotyping detection method based on nucleic acid tetrahedron probe modified printed gold electrode and kit
  • EGFR genotyping detection method based on nucleic acid tetrahedron probe modified printed gold electrode and kit
  • EGFR genotyping detection method based on nucleic acid tetrahedron probe modified printed gold electrode and kit

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Embodiment Construction

[0029] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0030] Such as Figure 1 to Figure 6 Shown, a kind of EGFR genotyping detection method based on nucleic acid tetrahedron probe modification printing gold electrode, comprises the following steps:

[0031] 1) Specific recognition and amplification of primers. In the reaction system, the concentration of the antisense primer chain is 200nM, the concentration of the sense primer chain is 20nM, and the thermal cycle conditions are 92°C for 15s, 58°C for 30s, 72°C for 30s, 45 cycles;

[0032] 2) Dissolve the four strands in TM hybridization buffer at equal concentrations, add TCEP at a final concentration of 30mM, heat the solution to 95°C for 5 minutes to fully untie the nucleic acid strands, then quickly transfer to 4°C for more than 30s, and the four nucleic acid strands The two are hybridized to form a tetrahedral structure, and the bottom thr...

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Abstract

The invention provides an EGFR genotyping (19del and L858R) detection method based on a nucleic acid tetrahedron probe modified printed gold electrode and a kit. The method comprises the following steps: identifying mutant genes in a genome sample by using different EGFR genotype specific primers labeled by biotin with different concentrations through LATE-PCR with an amplification retarding mutation system (ARMS); and the amplification capability of polymerase is utilized. The method has good detection sensitivity, can detect mutant genome DNA as low as 30 pg, has good detection specificity, and can detect 0.1% of mutant DNA under the interference of a large amount of non-mutant DNA. In addition, according to the method, the gene types of 19del and L858R of 13 non-small cell lung cancer patients are determined, and the results are consistent with commercial detection results.

Description

technical field [0001] The invention relates to an EGFR genotyping detection method and a kit based on a nucleic acid tetrahedron probe modified printed gold electrode. Background technique [0002] Non-small cell lung cancer (NSCLC) is one of the malignant tumors with the highest morbidity and mortality in clinical practice, and about 70% of the patients are already in the advanced stage when they are diagnosed. Compared with traditional chemotherapy, epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have less toxicity and better therapeutic effect, which can significantly increase the quality of life and overall survival of patients with advanced NSCLC. However, this approach can only be used for patients with sensitive mutations in the EGFR gene. Therefore, it is of great clinical significance to develop instant, simple, low-cost, high-sensitivity and high-specificity new methods for detecting EGFR gene mutations. [0003] In recent years, research...

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11G01N27/327
CPCC12Q1/6886G01N27/3275G01N27/3276C12Q2600/156
Inventor 王旭耀
Owner 王旭耀
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