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Method for detecting 21 genes of breast cancer in Asia

A gene detection, breast cancer technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of extensive classification methods, recurrence and metastasis, and limited classification accuracy.

Inactive Publication Date: 2018-09-25
上海美迪维康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this classification method is relatively extensive, and it is also found that the accuracy of this classification is limited in clinical practice, and a considerable number of patients who are considered to be at low risk of recurrence also have recurrence and metastasis in the later stage

Method used

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  • Method for detecting 21 genes of breast cancer in Asia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Cut 3 slices of 10um thick paraffin tissue slices, immediately put the slices into a 1.5ml centrifuge tube, add 160ul Deparaffinization Solution, vortex mix for 10s, briefly centrifuge, incubate at 56°C for 3min, cool at room temperature, add 150μl Buffer PKD, vortex mix , add 10 μl proteinase K to the lower layer of clear liquid, incubate at 56°C for 15 minutes, incubate at 80°C for 15 minutes, transfer the lower layer of colorless liquid to a new 2ml centrifuge tube. After cooling in ice bath and centrifugation, transfer the supernatant to a new centrifuge tube. Add 1 / 10 volume of DNase Booster Buffer and 10ul of DNase I stock solution, invert to mix, briefly centrifuge to collect the remaining liquid, incubate at room temperature, add Buffer RBC, ethanol, mix with a pipette tip, transfer 700ul of sample to RNeasy MinElute spin column , gently cap and centrifuge and discard to waste. Add 500ul Buffer RPE to the RNeasy MinElute spin column, gently cover the lid, 10000...

Embodiment 2

[0037] Example 2. Real-time fluorescent quantitative PCR technology detects gene expression

[0038] A total of 21 genes were selected as target genes, including 16 tumor-related genes and 5 reference genes. Tumor-related genes include: HER2-related genes include GRB7, ERBB2, estrogen-related genes include ESR1, PGR, BCL2, SCUBE2, proliferation-related genes include BIRC5, MKI67, CCNB1, MYBL2, AURKA, invasion-related genes include MMP11, CTSL2, and GSTM1, CD68, BAG1. Reference genes include ACTB, GAPDH, RPLPO, GUS, TFRC.

[0039] 1) RNA quality inspection

[0040] Use Agilent Bioanalyzer 2100 to evaluate the RNA quality, follow the operation manual to subpackage the Ladder in sequence, prepare the gel, balance, and detect the chip. After the completion of the gel run test, after the gel run is completed, take 350ul of DEPC water and add it to the electrode cleaner. Place the electrode cleaner on the detection platform for 10 seconds, take out the electrode cleaner, open the...

Embodiment 4

[0054] Example 4. Value-at-Risk Calculation

[0055] Firstly, the gene expression value is standardized so that the gene expression value is between 0 and 15, and the calculation formula is:

[0056] gene x =(x i -Min gene )*15 / (Max gene -Min gene )

[0057] gene x Indicates the value after normalization, x i Indicates the expression value measured by the experiment, Min gene is the minimum expression value of this gene in existing patients, Max gene is the maximum expression value of this gene in existing patients.

[0058] Calculate the recurrence risk value according to the above steps, and the calculation algorithm for each group is as follows:

[0059] HER2 group value = 0.9*GRB7+0.1*HER2, if the result is less than 8, set the score to 8;

[0060] Estrogen-related gene score = (0.8*ESR1+1.2*PGR+BCL2+SCUBE2) / 4;

[0061] Proliferation-related gene component value = (BIRC5+MKI67+MYBL2+CCNB1+AURKA) / 5, if the result is less than 6.5, set the score to 6.5;

[0062]...

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Abstract

The invention relates to the field of biotechnology, particularly to a method for detecting the 21 genes of breast cancer in Asia. According to the present invention, the frozen fresh cancer tissue samples are collected through the standard medical sample collection process, the expression levels of the selected 21 target genes are detected through the real-time fluorescence quantitative PCR technology, the risk valves are calculated according to the expression levels through the calculation formulas compiled based on different types of the genes, and evaluating is performed; and the precise method for detecting the 21 genes of breast cancer in Asia can be provided.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method for detecting Asian breast cancer 21 genes. Background technique [0002] Breast cancer is a common malignant tumor in women, which seriously threatens women's physical and mental health. Statistics show that breast cancer causes 458,000 deaths every year, and its incidence is on the rise worldwide. In recent years, the incidence of breast cancer in China has been rising rapidly and tends to be younger. Breast cancer has become the most common cancer among Chinese women, and the number of new breast cancer cases and deaths in China account for 12.2% and 9.6% of the world's annual breast cancer respectively. According to statistics, the five-year survival rate of breast cancer in the United States is 89%, and the five-year survival rate of breast cancer patients in China is 73.1%. With the awakening of people's awareness of early diagnosis of tumors, breast cancer screening has ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/6886
CPCC12Q1/6851C12Q2563/107C12Q2561/113
Inventor 刘雷侯文洪魏卫卫徐峰
Owner 上海美迪维康生物科技有限公司
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