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Mutant gene group for mammary cancer risk assessment and detection kit thereof

A technology for detecting kits and mutant genes, which is applied in the determination/inspection of microorganisms, recombinant DNA technology, DNA/RNA fragments, etc., which can solve the problems of method limitations and achieve high detection efficiency, high detection rate, and fast detection speed Effect

Inactive Publication Date: 2014-08-13
FUDAN UNIV SHANGHAI CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is that the current methods for assessing the risk of breast cancer are relatively limited, especially the research on the relationship between familial breast cancer susceptibility gene mutants and breast cancer risk in Asian and Chinese populations Very limited questions, providing a panel of mutated gene clusters for breast cancer risk assessment and their assay kits

Method used

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  • Mutant gene group for mammary cancer risk assessment and detection kit thereof
  • Mutant gene group for mammary cancer risk assessment and detection kit thereof
  • Mutant gene group for mammary cancer risk assessment and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of hereditary breast cancer susceptibility genes in Chinese population

[0045] Materials and methods:

[0046] From June 2011 to July 2012, among patients with breast cancer who were treated in the Cancer Hospital Affiliated to Fudan University, according to the following inclusion criteria, we performed continuous enrollment.

[0047] The inclusion criteria were: age of onset of breast cancer patients ≤35 years old, and at least one blood relative who was also a malignant tumor patient; age of onset of breast cancer >35 years old, but ≤50 years old, and at least two relatives of the same side (paternal lineage) or maternal line) blood relatives are also malignant tumors; breast cancer onset age > 50 years old, and at least 3 blood relatives on the same side (paternal or maternal line) are also malignant tumors.

[0048] The patient's medical history and family history were collected through questionnaires, and 5ml of peripheral blood was collected...

Embodiment 2

[0145] The primers needed for the synthetic kit of embodiment 2

[0146] Primer synthesis: The following primers were designed for the mutation sites described in the present invention, as shown in Table 7. For the primer sequences, please refer to SEQ ID NO: 1 to SEQ ID NO: 152 in the sequence listing.

[0147] Table 7 Tumor mutation site primers

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156] The meaning of Ex-Fw and Ex-Rv primers in Table 7 is the first pair of PCR primers in the nested PCR, and the meaning of In-Fw and In-Rv primers is the nested primers in the nested PCR reaction.

[0157] Utilize the primers shown in Table 7 to perform nested PCR on the patient's genomic DNA. The reaction system and reaction program of the nested PCR are conventional techniques in the art, and the nested PCR reaction program can be appropriately adjusted according to the size of the target fragment, and used The resulting PCR produc...

Embodiment 3

[0158] Example 3 Prepare the probes required for the detection kit

[0159] The capture target region of the gene is shown in Table 1: select the exon coding region of the gene to be tested and some flanking intron regions, and use the Agilent Sureselect XT Custom Kit (700Kb-34Mb) (the kit was purchased from Agilent Technologies, Inc. ) to design a probe, the sequence of the obtained probe is the same as or complementary to the gene sequence carrying the pathogenic mutation site shown in Table 4, please refer to the instruction manual of the kit for the design method of the probe.

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Abstract

The invention discloses a mutant gene group for mammary cancer risk assessment and a detection kit thereof. The mutant gene group is a mutant gene set comprising the following seven genes: BRCA1 gene, BRCA2 gene, TP53 gene, MLH1 gene, MLH3 gene, MSH3 gene and CDH1 gene. The mutant gene group of the seven genes totally carries 25 mutant sites disclosed as Table 3. The invention also discloses a detection kit of the mutant gene group for mammary cancer risk assessment, which comprises probes or primers for detecting the mutant genes in the mutant gene group. The detection kit disclosed by the invention has the advantages of high detection efficiency, high detection speed, high detectable rate and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a group of mutation gene groups for assessing the risk of breast cancer and a detection kit thereof. Background technique [0002] The most common cause of hereditary breast cancer is the mutation of BRCA1 and BRCA2 genes. It has been confirmed that early diagnosis and preventive intervention for mutation carriers can improve the survival rate of breast cancer. However, BRCA1 / 2 gene mutation can only explain a small part of the cause of hereditary breast cancer. Other highly dominant genes that play an important role in the genetic cause of breast cancer include TP53, PTEN and CDH1, etc. Genetic and hypodominant genes can also cause hereditary breast cancer. [0003] Clinically, in many tumor families with breast cancer as the proband, various malignant tumors are often combined, not just breast cancer and ovarian cancer. At the same time, the relationship between breas...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 胡震杨晓晨吴炅邵志敏
Owner FUDAN UNIV SHANGHAI CANCER CENT
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