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Human tumor specific BRAF mutation detection method

A tumor-specific, mutation detection technology, applied in the field of gene mutation detection, can solve the problems of long detection cycle and low detection sensitivity, and achieve the effect of high accuracy and high specificity

Pending Publication Date: 2022-07-29
上海睿璟生物科技有限公司
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Problems solved by technology

After decades of clinical application, it has been recognized by the majority of medical staff, but the detection sensitivity of this technology is relatively low, and the detection cycle is long

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  • Human tumor specific BRAF mutation detection method
  • Human tumor specific BRAF mutation detection method
  • Human tumor specific BRAF mutation detection method

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Embodiment Construction

[0022] The technical solution of the present patent will be described in further detail below in conjunction with specific embodiments.

[0023] see Figure 1-3 , a human tumor-specific BRAF mutation detection method, comprising a nucleic acid amplification reagent, the nucleic acid amplification reagent includes a BRAF V600E reaction solution and a BRAF V600K reaction solution, and the BRAF V600E reaction solution and the BRAF V600K reaction solution respectively contain for detection. Specific primers and probes for BRAF V600E and V600K mutations; the BRAFV600E reaction solution and the BRAF V600K reaction solution both contain a pair of upstream primers and downstream primers, and a TaqMan fluorescent probe. The primer probe sequences and modifications in the BRAF V600E reaction solution and the BRAF V600K reaction solution are as follows:

[0024] The upstream primer in the V600E reaction solution (sequence 1 in the sequence listing): 5'-GATTTTGGTCTAGCTACAGA-3'

[0025] ...

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Abstract

The invention discloses a human tumor specific BRAF mutation detection method in the technical field of gene detection, the detection method adopts a PCR kit for detecting human BRAF gene mutation, the PCR kit comprises a nucleic acid amplification reagent and a reference substance, the nucleic acid amplification reagent comprises a BRAFV600E reaction solution and a BRAFV600K reaction solution; the BRAFV600E reaction liquid and the BRAFV600K reaction liquid respectively contain a specific primer and a probe which are used for detecting the mutation of the BRAFV600E and the mutation of the V600K. The PCR kit for detecting human BRAF gene mutation adopts an allele specific amplification quantitative PCR technology to qualitatively detect BRAF gene V600E and V600K mutation, so that clinical diagnosis of colorectal cancer, thyroid cancer and melanoma is assisted, related patients are guided to treat and detect, and the PCR kit has high specificity, accuracy and high sensitivity.

Description

technical field [0001] The invention relates to a human tumor-specific BRAF mutation detection method in the technical field of gene mutation detection. Background technique [0002] BRAF gene is one of the most important human proto-oncogenes, located on human chromosome 7q34, composed of 18 exons, encoding a protein of about 94kDa. BRAF can activate the mitogen-activated protein kinase / extracellular signal-regulated kinase (MAPK / ERK) signaling pathway, which transmits the change information of various extracellular growth factors, cytokines and hormones to the nucleus, through ligands and casein. The amino acid kinase receptor surface binds to form dimers and autophosphorylates tyrosine residues. BRAF mutation directly leads to the activation of mitogen-activated extracellular signal-regulated kinase (MEK) and continues to activate ERK, resulting in the activation of the MAPK pathway, which in turn leads to uncontrolled cell cycle and circulation, and promotes tumorigenes...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/156C12Q2600/166C12Q2531/113C12Q2563/107
Inventor 石涵龚伟高伙妮
Owner 上海睿璟生物科技有限公司
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