Detecting system and kit for detecting BRAF gene mutation

A system and reaction system technology, applied in DNA/RNA fragmentation, recombinant DNA technology, microbial assay/inspection, etc., can solve the problems of insensitivity to treatment and low efficiency, improve sensitivity, reduce detection cost, and achieve absolute The effect of precise quantitative analysis

Pending Publication Date: 2018-11-16
上海赛安生物医药科技股份有限公司
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  • Claims
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Problems solved by technology

This suggests that the key detection of BRAF mutations in the future can provide guidance for the clinical treatment of colon cancer in the future. Patients with BRAF gene mutations have a low effective rate of EGFR-TKI drug treatment. Insensitivity to EGFR monoclonal antibody therapy

Method used

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  • Detecting system and kit for detecting BRAF gene mutation
  • Detecting system and kit for detecting BRAF gene mutation
  • Detecting system and kit for detecting BRAF gene mutation

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Embodiment Construction

[0040] The present invention is specifically described below through the examples, it is necessary to point out that the following examples are only used to further illustrate the present invention, and can not be interpreted as limiting the protection scope of the present invention, those skilled in the art can according to the above-mentioned present invention Contents Some non-essential improvements and adjustments are made to the present invention. In the following examples, unless otherwise specified, all reagents used are of analytical grade, and all reagents used can be obtained from commercial sources. For the experimental methods that do not indicate the specific conditions in the text, usually follow the conditions described in the book "Molecular Cloning Experiment Guide" published by Science Press, edited by J. Sambrook et al. in 2002, or according to the manufacturer's suggestion conditions of. Unless otherwise defined, all professional and scientific terms used ...

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Abstract

The invention discloses a detecting system and a kit for detecting BRAF gene mutation. The kit comprises digital PCR pre-mixed liquor and primer probe mixed liquor, wherein the primer probe mixed liquor comprises an upstream primer for detecting a site V600E of a 600rd codon, an downstream primer and a peptide nucleic acid probe; a nucleotide sequence of the upstream primer is as shown in SEQ ID No.1; a nucleotide sequence of the downstream primer is as shown in SEQ ID No.2; the nucleotide sequence of the peptide nucleic acid probe and the site V600E of the 600rd codon of the BRAF mutant geneare completely complementary, and the nucleotide sequence is as shown in SEQ ID No.3; the end 5 ' of the peptide nucleic acid probe is connected with a strong chelating group which consists of four amino acids. The detecting system and the kit for detecting BRAF gene mutation are high in sensitivity and specificity, are smaller in quantity demanded of samples, and can quickly, conveniently and accurately perform PCR quantitative detection.

Description

technical field [0001] The invention relates to a detection product for detecting BRAF gene mutation, which belongs to the field of biotechnology. Background technique [0002] The BRAF gene is located on chromosome 7q34 and is one of the most important proto-oncogenes in humans. It encodes a serine / threonine kinase and is an important transduction factor in the EGFR pathway RAS / RAF / MEK / MRK / MAPK and participates in the regulation of cell growth , differentiation and apoptosis and other physiological processes. Studies have shown that there are different rates of BRAF gene mutations in various malignant tumors such as malignant melanoma, lung cancer, and colorectal cancer, among which about 82% of the mutations are located at the 1799th nucleotide of exon 15, and the T mutation is A (T1799A ), causing the glutamic acid encoded by it to be replaced by valine (V600E). This mutation activates the BRAF protein and leads to the continuous activation of the downstream MEK-ERK sign...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/156C12Q2563/159C12Q2525/107C12Q2545/114
Inventor 王明赵国玲洪专
Owner 上海赛安生物医药科技股份有限公司
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