Breast cancer prognosis related gene mutation test kit and use method thereof

A kit and breast cancer technology, applied in the field of biomedicine, to achieve the effect of improved accuracy, rapid and effective early diagnosis

Active Publication Date: 2017-06-13
北京致成生物医学科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, there is no report on the application of the SNP sites of GFOD2, KCNK9 and MAP2 genes in the diagnosis and prognosis of breast cancer. If the SNPs related to the prognosis of breast cancer can be screened out as predictive indicators and the corresponding prognostic kits can be developed, it will be It has important guiding significance for the individualized treatment of breast cancer, and opens up a new way for its drug screening and efficacy evaluation

Method used

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  • Breast cancer prognosis related gene mutation test kit and use method thereof
  • Breast cancer prognosis related gene mutation test kit and use method thereof
  • Breast cancer prognosis related gene mutation test kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Extraction and Purification of Peripheral Blood DNA

[0058] 1. Sample collection

[0059] From January 2010 to December 2015, the inventor collected a large number of blood samples from patients with new breast cancer in Shenzhen Second People's Hospital. After sorting out the sample data, the inventor selected 25 samples that met the following criteria At the same time, 10 healthy women aged 25-55 were selected as controls for whole-exome microarray testing. The sample selection criteria were as follows:

[0060] (1) Breast cancer cases diagnosed by pathology, among which 3 patients have a family history of cancer and are marked as X1, X2, and X3 respectively;

[0061] (2) Have not received radiotherapy or chemotherapy before blood collection, and have no previous history of cancer;

[0062] (3) Healthy female controls matched with the age of the cases, and the demographic and clinical data of these samples were systematically collected.

[0063] 2. Extra...

Embodiment 2

[0073] The whole exome detection of embodiment 2 SNP

[0074] The two groups of people in Example 2 were detected by the whole exome chip to obtain relevant results.

[0075] 1. Library construction

[0076] Beijing Novogene Technology Co., Ltd. uses Agilent's liquid-phase chip capture system to efficiently enrich the DNA of the whole exon region of human beings, and then perform high-throughput and high-depth sequencing on the IlluminaHiseq platform. The Agilent SureSelectHumanAll ExonV5 kit was used for library construction and capture experiments, the reagents and consumables recommended in the instructions were strictly used, and the operation was performed according to the latest optimized experimental procedures.

[0077] The basic process of the experiment: Genomic DNA was randomly broken into fragments with a length of 180-280bp by a Covaris crusher, and after end repair and A-tailing, adapters were connected to both ends of the fragments to prepare a DNA library. Af...

Embodiment 3

[0088] Example 3 Using the risk scoring method to further analyze the risk of SNP and breast cancer

[0089] By comparing the genotype distribution frequencies of two groups of samples ("breast cancer case group" and "healthy female control group"), the inventors selected positively associated SNPs, and used the regression coefficient of a single SNP in the whole exome scanning sample as the weight , and further obtain the risk score, draw the ROC to evaluate the sensitivity and specificity of diagnosis, and then diagnose the ability of these SNPs to judge the onset of breast cancer. The joint analysis of all SNP markers found that the SNP mutation located at base 851 C / T of GFOD2 gene NM_030819:exon3, the SNP mutation of KCNK9 gene NM_001282534:exon2 base 803 G / T and the MAP2 gene NM_002374:exon7 The sensitivity and specificity of the SNP mutation of the 2996th base C / G are above 60%.

[0090] Thus, the inventors demonstrated that this locus marker was able to discriminate w...

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Abstract

The invention discloses a breast cancer prognosis related gene mutation test kit and a use method thereof. The test kit comprises reagents for testing susceptible SNP sites of GFOD2, KCNK9 and MAP2. The detailed information of the SNP sites is represented in table 1. The invention further discloses a biological reagent for testing the breast cancer prognosis related gene mutation. The test kit selects the application prospects in the breast cancer diagnosis and prognosis, of the SNP sites of three susceptible genes in the diagnosis of breast cancer, describes the influence brought by the SNP to the development of the breast cancer, and unveils the value of SNP in the diagnosis and prognosis. Therefore, the test kit and the biological reagent which is capable of using SNP gene to test the breast cancer prognosis is developed, the test kit not only can achieve the early period diagnosis and prognosis assessment, the accuracy is greatly increased, and the test kit provides therapeutic targets and an important basis for the clinical therapies such as gene therapy and medicine therapy.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a breast cancer prognosis-related gene mutation detection kit and a use method thereof. Background technique [0002] Breast cancer is a systemic disease, and its occurrence and development is a complex process involving multiple factors and links, including the activation of oncogenes and the inactivation of tumor suppressor genes. Therefore, gene mutation plays a very important role in the occurrence and development of breast cancer. [0003] Breast cancer is a multifactorial genetic variation disease, less than 10% is caused by a single gene defect. With the development of high-throughput gene technology, more and more breast cancer-related genes have been discovered, and potential genetic variations (single nucleotide polymorphisms and copy number variations) in these genes may cause differences in the therapeutic effects of breast cancer drugs . Due to the existence o...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 杨跃梅
Owner 北京致成生物医学科技有限公司
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