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Method for detecting HER 2 positivity of breast cancer through ddPCR technology

A detection object and detection kit technology, applied in the field of detecting HER2-positive breast cancer using ddPCR technology

Inactive Publication Date: 2016-10-05
SHANGHAI BIOTECAN PHARMA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In summary, there is still a lack of a method in the art that can accurately and effectively detect HER2 mutation positive

Method used

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  • Method for detecting HER 2 positivity of breast cancer through ddPCR technology
  • Method for detecting HER 2 positivity of breast cancer through ddPCR technology
  • Method for detecting HER 2 positivity of breast cancer through ddPCR technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0173] Example 1 Using ddPCR technology to detect breast cancer HER2 positive

[0174] (1) Extracting genomic DNA from tumor tissue of breast cancer patients;

[0175] Place the tissue sample on ice, cut the tissue sample the size of a grain of rice into a 1.5m1 centrifuge tube, and cut it as much as possible with a scalpel. Add 200 μl of buffer and 20 μl of proteinase K solution, and bathe in water at 56°C for more than 2 hours until the tissue is dissolved. Add 200 μl of buffer solution, mix thoroughly by inversion, and bathe in 70°C water for 10 minutes. Add 200 μl of absolute ethanol and shake fully for 15 seconds. Pour all the solution and precipitate into the adsorption column, centrifuge at 12000rpm for 30 seconds, and pour out the waste liquid in the collection tube. Add 500 μl of buffer solution to the adsorption column, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid in the collection tube. Add 700 μl of rinsing solution to the adsorption co...

Embodiment 2d

[0184] Embodiment 2ddPCR technology detects Herceptin drug resistance

[0185]No. 1-30 breast cancer patient samples were respectively used traditional IHC and FISH techniques and the method as described in Example 1 to measure the H1 / C1, H1 / E1, H1 / E2, H2 / C1, H2 / E1, H2 / E2 value. The results are shown in the table below:

[0186]

[0187]

[0188] Among them, patients 17 and 18 were tested for circulating tumor cell CTCs after their tumors were diagnosed, and 3 and 5 CTCs were detected, respectively, of which 2 and 3 CTCs were Her2-positive, respectively. Under the advice of the doctor, the patient started to use Herceptin after the operation, and there was no recurrence after one year, and the tumor was well controlled.

[0189] In addition, patients 13, 14, 15, 16, 19, and 20 were unable to be judged as positive for Her2 by IHC and FISH techniques, so doctors did not use Herceptin for them. Using the method of the present invention, patients No. 19 and No. 20 were i...

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Abstract

The invention provides a method for detecting HER 2 positivity of breast cancer through the ddPCR technology and specifically provides an HER 2 copy number variation detection kit. The kit is characterized by comprising two pairs of primers for detecting HER 2 genes and one or more pairs of primers for detecting reference genes of CEP 17 or EFTUD2 or a combination of CEP 17 and EFTUD2. By combining the ddPCR technology with a combination of multiple reference genes, whether a sample with suspended positivity determined through a conventional method has HER 2 positivity or not can be effectively judged, and therefore assistance and guide can be provided for breast cancer medication.

Description

technical field [0001] The invention relates to the technical field of molecular detection for guiding tumor treatment, in particular to a method for detecting breast cancer HER2 positive by using ddPCR technology. Background technique [0002] Breast cancer has become the malignant tumor with the highest incidence rate among women in the world, and it is one of the most common diseases that seriously affects women's physical and mental health and even threatens their lives. Patients with different molecular subtypes may have very different responses to clinical treatment and prognosis. [0003] Human epidermal growth factor receptor-2 (HER2) is a transmembrane glycoprotein with receptor tyrosine kinase (RTK) activity encoded by the HER2 (also known as ERBB2 or NEU) proto-oncogene, which can initiate tyrosine kinase Regulated signal transduction system. HER2 gene amplification and protein overexpression can overtransmit signals and stimulate cancer cell proliferation. HER...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张桢珍许骋易静丁飞飞徐本梅吴云鸣楼敬伟王剑鹏崔祥滨王臻王岭盛湲
Owner SHANGHAI BIOTECAN PHARMA
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