Breast cancer screening marker composition, selection method thereof and breast cancer screening kit

A breast cancer and composition technology, which is applied in the field of breast cancer screening kits, breast cancer screening marker compositions and its selection, can solve the problems of time-consuming, inaccurate detection, cumbersome breast cancer screening operations, etc. Effects of shortening time, reducing mortality, and improving inspection sensitivity

Pending Publication Date: 2022-01-28
杭州求臻医学检验实验室有限公司
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  • Abstract
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Problems solved by technology

[0003] The main purpose of the present invention is to provide a breast cancer screening marker composition, which aims to solve the problems of cumbersome, time-consuming and inaccurate detection of breast cancer screening in the prior art

Method used

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  • Breast cancer screening marker composition, selection method thereof and breast cancer screening kit
  • Breast cancer screening marker composition, selection method thereof and breast cancer screening kit
  • Breast cancer screening marker composition, selection method thereof and breast cancer screening kit

Examples

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Effect test

Embodiment

[0071] 1. Take 5mL of plasma, and use Qiagen plasma cell-free DNA extraction kit (Cat: 55204) to extract cfDNA.

[0072] 2. Take 20ng of extracted cfDNA and HinP1I (Cat: R0124, NEB), HpaII (Cat: R0171, NEB), AciI (Cat: R0551, NEB), HpyCH4IV (Cat: R0619, NEB) four methylation-sensitive endogenous Dicer (final concentration 10U / μL), 20 μL of the system, incubated at 37°C for 16h, inactivated at 80°C for 20min, and cut the background DNA.

[0073] 3. Use all the products after incubation as templates, add the above-mentioned 5 pairs of target-specific primers (50nM each) and 1 pair of internal reference primers (10nM), configure the system for multiplex PCR, and perform 8 cycles to obtain multiplex PCR products. The multiplex PCR reaction program is: 98°C / 45 9; 8cycle9 (98°C / 15 9, 55°C / 309, 72°C / 30 9)s 72°C / 1mins 4°C / hols.

[0074] 4. Take 1 μL of the multiplex PCR product in step 3 as a template, add the above 5 pairs of target-specific primers (0.25 μM each), and the correspon...

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Abstract

The invention provides a breast cancer screening marker composition, a selection method thereof and a breast cancer screening kit, and relates to the technical field of in-vitro diagnosis. According to the technical scheme, the breast cancer screening marker composition is hypermethylated in breast cancer and hypomethylated in white blood cells, and has a methylation sensitive incision enzyme digestion site; by utilizing the characteristic that methylation sensitive incision enzyme cannot cut methylation sites, non-tumor-source cfDNA is degraded, and tumor-source ctDNA is specifically amplified, so that the detection sensitivity is greatly improved; and the marking composition is small in quantity, simple to operate and short in time consumption. The breast cancer is detected and checked more accurately and effectively, so that the death rate of the breast cancer is reduced, and damage of the breast cancer to women is reduced.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a breast cancer screening marker composition, a selection method thereof, and a breast cancer screening kit. Background technique [0002] Breast cancer is the most common malignant tumor in women. Epidemiology shows that the prevalence of breast cancer is on the rise. It has become a common and frequently-occurring disease that seriously affects women's physical and mental health and even threatens their lives. In recent years, it has surpassed cervical cancer as the most common disease among women. The malignant tumor with the highest incidence rate is the number one killer of women's health. In the prior art, the 21-gene detection method is usually used for breast cancer detection and prognosis judgment. However, the 21-gene detection method involves many genes, and the operation is cumbersome, time-consuming, and the detection is not accurate enough. Contents of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 张怡然段小红杨春燕宋莹莹王冰王东亮
Owner 杭州求臻医学检验实验室有限公司
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