Noninvasive molecular subtyping kit and method for breast cancer

A technology for molecular typing and breast cancer, applied in biochemical equipment and methods, sequence analysis, instruments, etc., can solve the problems of judging prognosis and metastasis, result bias, and inability to obtain tissue samples, etc., and achieves simple sampling process and clinical Good experience and easy-to-obtain effect

Inactive Publication Date: 2019-08-09
广州市雄基生物信息技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This invasive diagnostic method increases patient suffering and the risk of tumor dissemination
In addition, due to the large heterogeneity of tumor tissues, the biopsy analysis of local punctures may cause biased results, and the pathological diagnosis of biopsy tissues also requires

Method used

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  • Noninvasive molecular subtyping kit and method for breast cancer
  • Noninvasive molecular subtyping kit and method for breast cancer
  • Noninvasive molecular subtyping kit and method for breast cancer

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Experimental program
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Embodiment 1

[0049] The described breast cancer non-invasive molecular typing product provided by the present invention comprises plasma free DNA extraction reagent (plasma free DNA extraction reagent is the DNA extraction kit of MagaBio company), library construction reagent (Ion Plus Fragment Library Kit reagent of Life technologies company) box), sequencing reagents, and sequencing chips. The library construction kit includes end repair, adapter ligation, and PCR amplification reagents.

[0050] It should be noted that the above plasma free DNA extraction reagents, library construction reagents and sequencing reagents can also be used from other manufacturers, such as:

[0051] Ion torrentproton sequencing platform: used sequencing reagents Ion PITMHi-QTMOT2200Kit article number: A26434 and Ion PITMHi-QTMSequencing 200Kit article number: A26433, sequencing chip Ion PITMChip Kitv3 article number: A26771.

[0052] Illumina Miseq sequencing platform: the sequencing reagent used MiSeq Reag...

Embodiment

[0054] Example Non-invasive molecular typing of breast cancer

[0055] Step 1: Plasma separation: Use two-step centrifugation for plasma separation (1) 4°C, 1600g centrifugation for 10 minutes to separate plasma from white blood cells, platelets, and red blood cells, and transfer the plasma to a 2mL centrifuge tube; (2) 4°C, 16000g Centrifuge for 10 min to remove residual cells, and transfer the supernatant to a 2 mL centrifuge tube to obtain peripheral blood plasma.

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Abstract

The invention provides a noninvasive molecular subtyping kit and method for a breast cancer and aims to provide a kit which is simple to use and high in detection accuracy and a noninvasive molecularsubtyping method for a breast cancer, which makes samples easy to obtain, is simple in sampling process, good in clinical experience, capable of achieving processized operation and very suitable for clinical application and non-disease diagnosis purposes. The technical scheme of the invention is that the noninvasive molecular subtyping kit for the breast cancer comprises a plasma free DNA extraction reagent, a library construction reagent, a sequencing reagent and a sequencing chip. The method is based on high-throughput sequencing data of plasma free DNA, nucleosome footprint difference information of a promoter region in the whole genome range in the plasma free DNA is obtained by means of bioinformatic analysis, and thus noninvasive molecular subtyping of four categories of a lumen A type, a lumen B type, an HER2 enrichment type and basal cell type of a breast cancer patient is achieved according to the difference of footprint information. The invention belongs to the technical field of biology.

Description

technical field [0001] The present invention provides a non-invasive molecular typing product of breast cancer, specifically, a non-invasive molecular typing product of breast cancer based on plasma cell-free DNA high-throughput sequencing technology, and the present invention also relates to a construction method of the molecular product, Belongs to the field of biotechnology. Background technique [0002] Breast cancer is the most common invasive cancer in women. According to the report of the World Health Organization in 2015, there are about 1.15 million new cases of breast cancer and about 410,000 deaths worldwide every year, and the incidence rate is increasing year by year. Breast cancer is also the most common cancer among Chinese women. Every year, the number of new breast cancer cases and deaths in China account for 12.2% and 9.6% of the world's total respectively. [0003] The molecular classification of breast cancer is closely related to the treatment and prog...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869G16B40/30G16B30/00
CPCC12Q1/6869G16B30/00G16B40/30C12Q2563/159C12Q2535/122C12Q2537/165
Inventor 李坤胥顺杨学习
Owner 广州市雄基生物信息技术有限公司
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