CN108410960A - A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes - Google Patents
A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes Download PDFInfo
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- CN108410960A CN108410960A CN201810439468.9A CN201810439468A CN108410960A CN 108410960 A CN108410960 A CN 108410960A CN 201810439468 A CN201810439468 A CN 201810439468A CN 108410960 A CN108410960 A CN 108410960A
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Abstract
The exon primer for providing a pair of of BRCA2 genes of the present invention includes the exon reverse primer of the exon forward primer of BRCA2 genes and BRCA2 genes, and the exon forward primer of BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.The present invention also provides a kind of kit of the exon primer comprising a pair of BRCA2 genes, which can be used for breast cancer detection.Kit provided by the invention can detection BRCA2 gene mutations quick, accurate, easy to operate and effectively avoiding pollution kit, solve and carry out that BRCA2 detection in Gene Mutation programs are cumbersome, somewhat expensive or the problems such as time-consuming, accuracy is low and pollution in the prior art.
Description
Technical field
The present invention relates to biological gene detection technique field, more particularly to the exon primer and breast of a kind of BRCA2 genes
Cancer oncogene detection kit.
Background technology
Female mammary gland is made of skin, fibr tissue, corpus mamma and fat, and breast cancer is happened on mammary gland gland
The malignant tumour of skin tissue.99% is happened at women in breast cancer, and male only accounts for 1%.
Mammary gland is not the vitals for maintaining human life activity, and breast cancer in situ is simultaneously not fatal；But due to breast cancer
The cell loss characteristic of normal cell, connects between cell loose, is easy to fall off.Cancer cell once falls off, and free cancer is thin
Born of the same parents can send out whole body with blood or lymph, form transfer, threat to life.Breast cancer, which has become, at present threatens women body and mind strong
The kinds of tumor of health.
Global breast cancer incidence is in rising trend always since the late 1970s.8, U.S. women is in life
1 people is just had to suffer from breast cancer.China is not the country occurred frequently of breast cancer, but unsuitable optimistic, in recent years China's breast cancer incidence
Growth rate is but higher by national 1~2 percentage point occurred frequently.According to National Cancer Center and prevention and control of diseases office of the Ministry of Public Health 2012
The pathogenesis of breast carcinoma data in 2009 announced are shown：National tumour registration area breast cancer incidence occupies female malignant
1st, female mammary gland cancer morbidity (rough and careless) whole nation adds up to 42.55/10 ten thousand, and city is 51.91/10 ten thousand, and rural area is
23.12/10 ten thousand.
Human breast carcinoma tumor susceptibility gene 2 (breast cancer 2, early onset), is abbreviated as BRCA2.BRCA2 is
The tumor susceptibility gene of breast cancer and oophoroma with genetic predisposition, plays an important role in terms of DNA damage reparation.Breast cancer
Occurrence and development are series of genes, the coefficient result of environmental factor.Numerous epidemiological study prompts, menstruation, reproduction, life
The generation of the factors such as mode (such as first menses is early, in menopause evening, gestation, has an abortion, takes oral contraceptive) living and breast cancer is also close
Correlation, but when individual of different genes background is in same exposure level the neurological susceptibility of breast cancer is different.This gene
The difference of background determines the size of the individual risk that suffers from breast cancer.BRCA2 be find after the BRCA1 familial breast cancer/
Ovarian cancer susceptible gene, coded product are a kind of tumor suppressor.This protective protein is adjusted by homologous recombination and part
The activity for saving Rad51 protein factors participates in the reparation of double-strand DNA damage, to prevent the growth and development of cancer cell.Study table
Bright, the BRCA2 genes of mutation can cause the occurrence risk of breast cancer, oophoroma and cancer of pancreas to increase.
Invention content
Present invention aims at the exon primer for providing a kind of BRCA2 genes of quick, accurate, easy to operate detection and
Mastocarcinoma gene detection kit solves and carries out that BRCA2 detection in Gene Mutation programs are cumbersome, somewhat expensive in the prior art
Or the problems such as time-consuming, accuracy is low and pollutes.
According to an aspect of the present invention, the exon primer of a pair of of BRCA2 genes is provided, including BRCA2 genes is outer
The exon forward primer of the exon reverse primer of aobvious sub- forward primer and BRCA2 genes, BRCA2 genes is SEQ No.1：
CAGCATGTCACCCAG；The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
According to an aspect of the present invention, a kind of mastocarcinoma gene detection kit is provided, kit includes a pair of BRCA2
The exon primer of gene, the exon primers of a pair of of BRCA2 genes include BRCA2 genes exon forward primer and
The exon forward primer of the exon reverse primer of BRCA2 genes, BRCA2 genes is SEQ No.1：
CAGCATGTCACCCAG；The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
According to another aspect of the present invention, a kind of mastocarcinoma gene detection kit is provided, kit further includes PCR slow
Fliud flushing, dNTP mixed liquors, fluorescent dye, MgCl2, TqqDNA polymerase.
According to another aspect of the present invention, a kind of mastocarcinoma gene detection kit, fluorescent dye LC are provided
Green Plus dyestuffs.
According to another aspect of the present invention, a kind of mastocarcinoma gene detection kit is provided, dNTP mixed liquors include
The mixed liquor of 10mM dATP, 10mM dCTP, 10mM dTTP, 10mM dGTP.
According to another aspect of the present invention, a kind of mastocarcinoma gene detection kit is provided, PCR buffer solutions include
Triscl, potassium chloride, magnesium chloride.
The purpose of the present invention is to provide pair of primers and application primer as PCR reaction kits, and kit can be used
Mastocarcinoma gene diagnose, using detection BRCA gene mutations the phenomenon that, by testing result be breast cancer judgement be used as according to
According to.
Specific implementation mode
The technical solution that the present invention is further explained With reference to embodiment.
Following embodiment is intended to illustrate invention rather than limitation of the invention further.
The present invention provides a kind of mastocarcinoma gene detection kit, and kit includes that the exon of a pair of of BRCA2 genes draws
Object, the exon primer of a pair of of BRCA2 genes include the exon of the exon forward primer and BRCA2 genes of BRCA2 genes
Reverse primer.
The exon forward primer of BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
Mastocarcinoma gene detection kit further includes PCR buffer solutions, dNTP mixed liquors, fluorescent dye, MgCl2, TqqDNA
Polymerase；Fluorescent dye is LC Green Plus dyestuffs.
Wherein dNTP mixed liquors include the mixed liquor of 10mM dATP, 10mM dCTP, 10mM dTTP, 10mM dGTP.PCR
Buffer solution includes Triscl, potassium chloride, magnesium chloride.
Embodiment 1：
1. extracting DNA profiling：For familial breast cancer, patient is detected, paraffin section tissue BRCA2 abrupt climatic change men
The paraffin section of race heredity patient with breast cancer 10 extracts DNA as template using commercial kits.
Use instrument：LlightCycler
480 quantitative fluorescent PCR instruments of TM are analyzed
2：It is formulated as follows reaction system
Primer sequence is provided：
The exon forward primer of BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
Form PCR reaction systems：
| Ingredient (concentration) | The volume of addition |
| 10×PCR Buffer | 2.0μL |
| 5μm MgCl2 | 0.5μL |
| 10 μm of (m/mol) dNTP mixed liquors | 0.5μL |
| 10 μm of LC Green Plus dyestuffs | 1.0μL |
| 10 μm of primers | 1.0μL |
| 5 μm of TqqDNA polymerases | 0.2μL |
| 10 μm of templates | 1.0μL |
| ddH2O | 13.8μL |
Wherein dNTP mixed liquors include the mixed liquor of 10mM dATP, 10mM dCTP, 10mM dTTP, 10mM dGTP.PCR
Buffer solution includes Triscl, potassium chloride, magnesium chloride.
3.PCR-HRM reaction conditions：
4. 11 exons of 10 familial breast cancer patient's BRCA2 genes of testing result, have 3 examples to detect BRCA2 bases
11 exons mutations of cause, it is seen that the present invention has good reference significance to breast cancer diagnosis.
Embodiment 2：
Choose familial breast cancer patient same as Example 1, patient with breast cancer's paraffin section tissue BRCA2 abrupt climatic changes
The paraffin section of familial inheritance patient with breast cancer 10 extracts DNA as template using commercial kits.
Use instrument：LlightCycler
480 quantitative fluorescent PCR instruments of TM are analyzed
Primer sequence：
The exon forward primer of BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
PCR reaction systems：
Wherein dNTP mixed liquors include the mixed liquor of 10mM dATP, 10mM dCTP, 10mM dTTP, 10mM dGTP.PCR
Buffer solution includes Triscl, potassium chloride, magnesium chloride.
PCR-HRM reaction conditions：
Identical as embodiment one, the exon of 10 familial breast cancer patient's BRCA2 genes of testing result has the inspection of 3 examples
Measure the exons mutation of BRCA2 genes, it is seen that the present invention has good reference significance to breast cancer diagnosis.
Embodiment 3：
Choose familial breast cancer patient same as Example 1, patient with breast cancer's paraffin section tissue BRCA2 abrupt climatic changes
The paraffin section of familial inheritance patient with breast cancer 10 extracts DNA as template using commercial kits.
Use instrument：LlightCycler
480 quantitative fluorescent PCR instruments of TM are analyzed
Primer sequence：
The exon forward primer of BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of BRCA2 genes：GTTTATCACCTGTGTCT.
PCR reaction systems：
PCR-HRM reaction conditions：
Identical as embodiment one, the exon of 10 familial breast cancer patient's BRCA2 genes of testing result has the inspection of 3 examples
Measure the exons mutation of BRCA2 genes, it is seen that the present invention has good reference significance to breast cancer diagnosis.
The foregoing examples are merely illustrative of the technical concept and features of the invention, its object is to allow the person skilled in the art to be
It cans understand the content of the present invention and implement it accordingly, it is not intended to limit the scope of the present invention.It is all smart according to the present invention
The equivalent transformation or modification that refreshing essence is done, should be covered by the protection scope of the present invention.
Sequence table
<110>The Suzhou bio tech ltd Hai Miao
<120>A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 15
<212> DNA
<213>（Artificial sequence）
<400> 1
cagcatgtca cccag 15
<210> 2
<211> 17
<212> DNA
<213>（Artificial sequence）
<400> 2
gtttatcacc tgtgtct 17
Claims (6)
1. a kind of exon primer of BRCA2 genes, it is characterised in that：Exon forward primer including BRCA2 genes and
The exon reverse primer of BRCA2 genes,
The exon forward primer of the BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of the BRCA2 genes：GTTTATCACCTGTGTCT.
2. a kind of mastocarcinoma gene detection kit, it is characterised in that：The kit includes the exon of a pair of of BRCA2 genes
The exon primer of primer, the pair of BRCA2 genes includes the exon forward primer and BRCA2 genes of BRCA2 genes
Exon reverse primer,
The exon forward primer of the BRCA2 genes is SEQ No.1：CAGCATGTCACCCAG；
The exon reverse primer SEQ No.2 of the BRCA2 genes：GTTTATCACCTGTGTCT.
3. a kind of mastocarcinoma gene detection kit according to claim 2, it is characterised in that：The kit further includes
DNTP mixed liquors, fluorescent dye, MgCl2, TqqDNA polymerases.
4. a kind of mastocarcinoma gene detection kit according to claim 3, it is characterised in that：The fluorescent dye is LC
Green Plus dyestuffs.
5. a kind of mastocarcinoma gene detection kit according to claim 4, the dNTP mixed liquors include 10mM dATP,
The mixed liquor of 10mM dCTP, 10mM dTTP, 10mM dGTP.
6. a kind of mastocarcinoma gene detection kit according to claim 5, the PCR buffer solutions include Triscl, chlorine
Change potassium, magnesium chloride.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810439468.9A CN108410960A (en) | 2018-05-09 | 2018-05-09 | A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810439468.9A CN108410960A (en) | 2018-05-09 | 2018-05-09 | A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes |
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| CN108410960A true CN108410960A (en) | 2018-08-17 |
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| CN201810439468.9A Pending CN108410960A (en) | 2018-05-09 | 2018-05-09 | A kind of the exon primer and mastocarcinoma gene detection kit of BRCA2 genes |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921831A (en) * | 2010-03-29 | 2010-12-22 | 苏州工业园区为真生物医药科技有限公司 | Rapid detection of BRCA (Breast Cancer) genic mutation |
| CN204097454U (en) * | 2014-09-12 | 2015-01-14 | 上海中优医药高科技有限公司 | Familial breast cancer BRCA1 and BRCA2 gene detecting kit |
| CN106929564A (en) * | 2015-12-30 | 2017-07-07 | 安诺优达基因科技(北京)有限公司 | breast cancer susceptibility gene detection kit |
| CN107523609A (en) * | 2016-06-22 | 2017-12-29 | 海门中科基因生物科技有限公司 | Hereditary breast cancer and oophoroma BRCA1/2 gene hot mutant site detection kits |
| CN107841558A (en) * | 2017-09-25 | 2018-03-27 | 北京青航基因科技有限公司 | Specific primer and application for the mutation of rapid screening BRCA1/2 gene extrons |
-
2018
- 2018-05-09 CN CN201810439468.9A patent/CN108410960A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921831A (en) * | 2010-03-29 | 2010-12-22 | 苏州工业园区为真生物医药科技有限公司 | Rapid detection of BRCA (Breast Cancer) genic mutation |
| CN204097454U (en) * | 2014-09-12 | 2015-01-14 | 上海中优医药高科技有限公司 | Familial breast cancer BRCA1 and BRCA2 gene detecting kit |
| CN106929564A (en) * | 2015-12-30 | 2017-07-07 | 安诺优达基因科技(北京)有限公司 | breast cancer susceptibility gene detection kit |
| CN107523609A (en) * | 2016-06-22 | 2017-12-29 | 海门中科基因生物科技有限公司 | Hereditary breast cancer and oophoroma BRCA1/2 gene hot mutant site detection kits |
| CN107841558A (en) * | 2017-09-25 | 2018-03-27 | 北京青航基因科技有限公司 | Specific primer and application for the mutation of rapid screening BRCA1/2 gene extrons |
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