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Triple-negative breast cancer metastasis-associated susceptibility gene tnf-α mutation detection kit and detection method

A technology of triple-negative breast cancer and a detection kit, which is applied in the fields of molecular biology and genetic diagnosis, and can solve the problems of poor prognosis, high risk of triple-negative breast cancer metastasis, and no molecular biology and genetic diagnosis methods for TNBC patients , to achieve the effect of ingenious design, accurate and reliable results

Active Publication Date: 2016-08-31
SHANDONG TUMOR HOSPITAL
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Problems solved by technology

Although triple-negative breast cancer has a high risk of metastasis and poor prognosis, there are currently no molecular biological and genetic diagnostic methods that can assess the risk of metastasis in TNBC patients

Method used

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  • Triple-negative breast cancer metastasis-associated susceptibility gene tnf-α mutation detection kit and detection method
  • Triple-negative breast cancer metastasis-associated susceptibility gene tnf-α mutation detection kit and detection method

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Experimental program
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Effect test

Embodiment 1

[0033] A triple-negative breast cancer metastasis-related susceptibility gene TNF-α mutation detection kit, its components and contents are as follows: including 1.0ul of 10×PCR reaction buffer, 25mM MgCl 2 Solution 0.5ul, dNTP Mixture (2.5mM) 1.0ul, TaqDNA polymerase 0.25ul (5U / ul), specific primers F and R (10uM) each 0.25ul, sterilized distilled water 6.0ul. This kit is used for one person-time detection, and the storage temperature is -20°C.

[0034] Step 1: DNA extraction from peripheral blood of patients with triple-negative breast cancer:

[0035] Genomic DNA from peripheral blood of patients was extracted using a blood genomic DNA extraction system (spin column method).

[0036] Step 2: PCR reaction, copy the target fragment:

[0037] (1) Design specific primers according to the gene sequence of the TNF-α promoter region to amplify the DNA fragment containing the rs1800629 site. The designed primer sequences are as follows:

[0038] F: 5’-CCCCAAAAGAAATGGAGGCAATAGGTT...

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Abstract

The invention provides a detection kit for triple negative breast cancer metastasis susceptibility gene TNF-alpha mutation. The detection kit comprises a specific primer and a related reaction solution. Meanwhile, the invention provides a detection method of the detection kit. The detection kit is ingenious in design, simple, feasible and capable of being popularized in hospitals at different levels, helps to evaluate the tumor metastasis risk of a TNBC patient and is beneficial for early intervening and treating the patient clinically, and results are accurate and reliable.

Description

technical field [0001] The invention relates to the fields of molecular biology and gene diagnosis, and is a kit for detecting the mutation of the susceptibility gene TNF-α related to the metastasis of triple-negative breast cancer, which is predicted by detecting the 308G>A genetic variation (rs1800629) in the promoter region of the TNF-α gene Metastatic risk in triple-negative breast cancer. Background technique [0002] Triple-negative breast cancer (TNBC), as a special subtype, specifically refers to estrogen receptor (estrogen receptor, ER), progesterone receptor (progesterone receptor, PR), epidermal growth factor receptor 2 (human epidermal growth factor receptor 2, HER2) negative breast cancer, accounting for about 15% of all breast cancers, is more likely to occur in young women, and about 1 / 3 of the patients have recurrence and metastasis within 2-3 years after surgery, and Rapid progression to death after relapse. Early assessment of the risk of metastasis in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/156
Inventor 李慧慧孟雪宋宝翟侃常春晓刘聪徐娜商玉红
Owner SHANDONG TUMOR HOSPITAL
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