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Gene combination for instructing individualized treatment by medicines such as geftinat and Tarceva

A gene combination and drug technology, applied in the field of genes, can solve problems such as delaying the timing of treatment, failure to achieve treatment goals, and complicated procedures

Inactive Publication Date: 2011-08-10
SHANGHAI BIOTECAN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Defects of existing clinical medication methods: In the treatment of many diseases, improper medication often leads to side effects or failure to achieve therapeutic goals due to insufficient dosage
This process of adjusting medication is complicated, time-consuming, and delays the timing of treatment

Method used

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  • Gene combination for instructing individualized treatment by medicines such as geftinat and Tarceva
  • Gene combination for instructing individualized treatment by medicines such as geftinat and Tarceva
  • Gene combination for instructing individualized treatment by medicines such as geftinat and Tarceva

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 A method for identifying EGFR-Exon19 mutations

[0031] 1. Extract the genomic DNA of the patient's tumor tissue;

[0032] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer ( Contains Mg 2+ ) 2 μl, and the rest was sterile distilled water; the PCR reaction conditions were: denaturation at 95°C for 2 min, followed by denaturation at 95°C for 30 sec, annealing at 55°C for 30 sec, extension at 72°C for 1 min, and 45 cycles, and finally extension at 72°C for 7 min, and storage at 4°C;

[0033] 3. Gel electrophoresis analysis of PCR amplification products:

[0034] a. Rinse the electrophoresis tank and comb with distilled water. Put it on a level table and set up the comb. 1.5% agarose can be prepared for electrophoresis.

[0035] b. Put 50ml of 1×TAE electrophoresis ...

Embodiment 2

[0054] Example 2 A method for identifying EGFR-Exon21 mutations

[0055] 1. Extract the genomic DNA of the patient's tumor tissue;

[0056] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer ( Contains Mg 2+ ) 2 μl, and the rest was sterile distilled water; the PCR reaction conditions were: denaturation at 95°C for 2 min, followed by denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, extension at 72°C for 1 min, and 45 cycles, and finally extension at 72°C for 7 min, and storage at 4°C;

[0057] 3. Gel electrophoresis analysis of the PCR amplification product: the specific steps are the same as in Example 1.

[0058] The result is as figure 1 As shown, band 4 represents EGFR-Exon21, and its PCR product is 247bp.

[0059] 4. Sequencing of PCR amplification products: the ...

Embodiment 3

[0061] Example 3 A method for identifying Kras-codon12 / 13 mutations

[0062] 1. Extract the genomic DNA of the patient's tumor tissue;

[0063] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer ( Contains Mg 2+ ) 2 μl, the rest was sterile distilled water; the PCR reaction conditions were: denaturation at 95°C for 2 minutes, followed by denaturation at 95°C for 30 sec, annealing at 50°C for 30 sec, extension at 72°C for 1 min, and 45 cycles, and finally extension at 72°C for 7 min, and storage at 4°C;

[0064] 3. Gel electrophoresis analysis of the PCR amplification product: the specific steps are the same as in Example 1.

[0065] The result is as figure 1 As shown, band 1 represents Kras-codon12 / 13, and its PCR product is 235bp.

[0066] 4. Sequencing of PCR amplification produc...

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Abstract

The invention belongs to the technical field of gene techniques and provides a gene combination for instructing the individualized treatment by medicines such as geftinat and Tarceva, which comprises epidermal growth factor receptor (EGFR)-Exon19, EGFR-exon21, Kras-codon12 / 13, and Kras-codon61. In the invention, instruction on medicine allergy is given to a patient to be treated by EGFR-tyrosine kinase inhibitors (TKI) such as geftinat and Tarceva by detecting the mutation condition of the gene loci, namely the EGFR-Exon19, EGFR-exon21, Kras-codon12 / 13, and Kras-codon61, in the tumor tissue of the patient, so that the treatment effect is maximized and the side effect is reduced to the lowest degree.

Description

technical field [0001] The invention belongs to the field of gene technology, and relates to a group of gene combinations used to guide the individualized treatment of Iressa and Tarceva drugs. Background technique [0002] The epidermal growth factor receptor (EGFR) tyrosine kinase-mediated cell growth signaling pathway plays an important role in the formation and progression of cancer. EGFR is overexpressed in patients with many different solid tumors, including non-small cell lung cancer (NSCLC), breast cancer. Ovarian cancer, head and neck cancer, gastric cancer, prostate cancer, bladder cancer, colon cancer and glioma, etc., are often associated with poor prognosis. Chemoradiotherapy resistance, tumor angiogenesis and tumor metastasis are related, and have become ideal targets for tumor therapy. In recent years, many new anti-tumor drugs targeting this target have been developed one after another, and have achieved encouraging curative effects in the treatment of vari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 楼敬伟黄慧徐玲丽
Owner SHANGHAI BIOTECAN PHARMA
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