CN107574236A - It is a kind of that the method for assessing Alzheimer's disease risk is sequenced based on sanger - Google Patents
It is a kind of that the method for assessing Alzheimer's disease risk is sequenced based on sanger Download PDFInfo
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- CN107574236A CN107574236A CN201610537027.3A CN201610537027A CN107574236A CN 107574236 A CN107574236 A CN 107574236A CN 201610537027 A CN201610537027 A CN 201610537027A CN 107574236 A CN107574236 A CN 107574236A
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- alzheimer
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Abstract
The present invention relates to molecular biology and field of bioinformatics, is a kind of method for assessing Alzheimer's disease risk based on sanger sequencings (also known as generation sequencing).It is characterized in that by the way that Alzheimer's disease related gene APOE is sequenced, the genotype in site related to Alzheimer's disease on the gene is determined, with reference to bioinformatics, genetic background support is provided to assess Alzheimer's disease risk.The present invention can simply, quickly and easily qualitative evaluation Alzheimer's disease risk, so as to accomplish disease early know, early prevention.
Description
Technical field
This method is related to a kind of molecular biology and field of bioinformatics, is that a kind of sanger that is based on is surveyed more specifically
The method that sequence assesses Alzheimer's disease risk.
Background technology
Senile dementia is a kind of central nervous system degeneration, shows as the decline of elaborative faculty and intelligence, all finally
Daily self care ability, general 65 years old sequela can be lost.Currently without effective treatment method, but researcher and
Doctor is understanding its pathogenesis, to strive for improving disease symptomses and finally slow down or even prevent the development of illness.Lose
Biography factor is bigger than acquired disposition to senile dementia role, it is known that gene such as APOE gene diversities can shadow
The incidence of disease is rung, and such as influence of hypertension, high fat of blood, diabetes etc. for Alzheimer's disease of common acquired disposition is not so good as
Its influence of APOE gene pairs is big.
The content of the invention
The purpose of the present invention aim to provide it is a kind of can simple, quickly and easily qualitative evaluation Alzheimer's disease illness wind
Danger, so as to so that High risk group prevention ahead of time, avoids the generation of Alzheimer's disease.
To reach above-mentioned purpose, specific design of the invention is：The genotype of different people is different, and different genotype with
Alzheimer's disease incidence probability difference has relation.This patent obtains different people base using sanger sequencings (generation sequencing)
Because of type, so as to judge the ill probability for suffering from Alzheimer's disease, reach the early purpose known, early prevented.
Based on above-mentioned design, the present invention adopts the following technical scheme that：
(1) DNA extractions are carried out to coming from human mouth mucosa cells or other cells
(2) performing PCR (PCR) amplification is entered to the DNA extracted, its relevant primer is as follows, and the primer will not
Cause the amplification of non-specific band：
I.Rs429358 sites
Forward 5′-CTACAAATCGGAACTGGAGGAACA-3′
Reverse 5′-AGCCGCTTACGCAGCTT-3′
Ii.Rs7412 sites
Forward 5′-CCTCCCACCTGCGCAAG-3′
Reverse 5′-CGGCCCTGTTCCACCAG-3′
(3) sanger sequencings are carried out to corresponding amplimer
(4) sequencing result is analyzed, the true base composition of aim sequence is obtained, the different genes in each site can be obtained
Type.Remarks：There are tri- kinds of CC, CT, TT in rs429358 sites, and there is tri- kinds of CC, CT, TT in rs7412 sites.
(5) genotype in the rs429358 and rs7412 sites obtained according to sequencing judges the ill wind of Alzheimer's disease
Danger.
| Risk | rs429358 | rs7412 |
| It is low | TT | TT |
| It is low | TT | CT |
| Typically | TT | CC |
| It is high | CC | CC |
| It is high | CC | CT |
| Typically | CC | TT |
| It is high | CT | CC |
| It is high | CT | CT |
| It is low | CT | TT |
The present invention uses above-mentioned technical proposal, has the advantages that：
(1) present invention is using sanger sequencings (generation sequencing), and sequence measurement is simple and easy, safe and reliable, economical and efficient；
(2) present invention enters performing PCR (polymerization to the DNA extracted using Rs429358 sites and Rs7412 sites as primer
PCR) amplification, and amplified production is sequenced by sanger sequencings, it can quickly, scientifically judge to suffer from A Er
The ill probability of Ci Haimo diseases；
(3) present invention is based on sanger sequencing technologies, can overcome biography with qualitative evaluation Alzheimer's disease risk
For system based on the deficiency in cognitive function slump evaluations, providing a kind of science for science judge Alzheimer's disease risk can
Capable solution.
Brief description of the drawings
Fig. 1 is that the method flow diagram for assessing Alzheimer's disease risk is sequenced based on sanger by a kind of of the present invention；
Embodiment
With reference to specific embodiment, the present invention is furture elucidated, it should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention, after the present invention has been read, various equivalences of the those skilled in the art to the present invention
The modification of form falls within the application appended claims limited range.
As shown in figure 1, a kind of the method for assessing Alzheimer's disease risk is sequenced based on sanger, following step is included
Suddenly：
S-1 takes human mouth mucosa cells or other cells；
STb gene in S-2 extraction cells；
S-3 is based on Rs429358 sites and Rs7412 sites primer, enters performing PCR (polymerase chain to the target DNA extracted
Formula is reacted) amplification；Wherein,
Rs429358 sites primer：
Forward 5′-CTACAAATCGGAACTGGAGGAACA-3′
Reverse 5′-AGCCGCTTACGCAGCTT-3′
Rs7412 sites primer：
Forward 5′-CCTCCCACCTGCGCAAG-3′
Reverse 5′-CGGCCCTGTTCCACCAG-3′
S-4 is sequenced using sanger and S-3 amplified production is sequenced；
S-5 analyzes sequencing result, obtains the different genotype in each site；Wherein,
PS1 sites：There is tri- kinds of CC, CT, TT in rs429358 sites；
PS2 sites：There is tri- kinds of CC, CT, TT in rs7412 sites.
S-6 qualitative evaluation Alzheimer's disease risks, are specifically included as follows：
S-6-1 assumes that rs429358 site informations are TT, and rs7412 site informations are TT, then Alzheimer's disease trouble
Sick risk assessment is low；
S-6-2 assumes that rs429358 site informations are TT, and rs7412 site informations are CT, then Alzheimer's disease trouble
Sick risk assessment is low；
S-6-3 assumes that rs429358 site informations are TT, and rs7412 site informations are CC, then Alzheimer's disease trouble
Sick risk assessment is general；
S-6-4 assumes that rs429358 site informations are CC, and rs7412 site informations are CC, then Alzheimer's disease trouble
Sick risk assessment is height；
S-6-5 assumes that rs429358 site informations are CC, and rs7412 site informations are CT, then Alzheimer's disease trouble
Sick risk assessment is height；
S-6-6 assumes that rs429358 site informations are CC, and rs7412 site informations are TT, then Alzheimer's disease trouble
Sick risk assessment is general；
S-6-7 assumes that rs429358 site informations are CT, and rs7412 site informations are CC, then Alzheimer's disease trouble
Sick risk assessment is height；
S-6-8 assumes that rs429358 site informations are CT, and rs7412 site informations are CT, then Alzheimer's disease trouble
Sick risk assessment is height；
S-6-9 assumes that rs429358 site informations are CT, and rs7412 site informations are TT, then Alzheimer's disease trouble
Sick risk assessment is low.
Claims (3)
- A kind of 1. method for assessing Alzheimer's disease risk, it is characterised in that obtained according to sanger sequencings (generation is sequenced) It is the genotype in two sites of rs429358 and rs7412 of APOE genes to the gene related to Alzheimer's disease, it is qualitative to comment Estimate Alzheimer's disease risk.
- 2. a kind of method that assessment Alzheimer's disease risk is sequenced based on sanger according to claim 1, its It is characterised by, specifically the step of assessment is：A) to coming from human mouth mucosa cells or other cells using the DNA in DNA extraction kit extraction total cell.B) performing PCR (PCR) amplification is entered to the DNA extracted, its relevant primer is as follows：I.Rs429358 sitesForward 5′-CTACAAATCGGAACTGGAGGAACA-3′Reverse 5′-AGCCGCTTACGCAGCTT-3′Ii.Rs7412 sitesForward 5′-CCTCCCACCTGCGCAAG-3′Reverse 5′-CGGCCCTGTTCCACCAG-3′C) sanger sequencings are carried out to corresponding amplimerD) sequencing result is analyzed, the different genotype in each site can be obtained.Remarks：There is CC, CT, TT tri- in rs429358 sites Kind, there is tri- kinds of CC, CT, TT in rs7412 sites.E) genotype in the rs429358 and rs7412 sites obtained according to sequencing judges the risk of Alzheimer's disease.
- 3. the method for the assessment Alzheimer's disease risk according to claim 1 and 2, it is characterised in that everyone Genotype can be covered by above-mentioned analysis result, and obtain the risk of Alzheimer's disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610537027.3A CN107574236A (en) | 2016-07-02 | 2016-07-02 | It is a kind of that the method for assessing Alzheimer's disease risk is sequenced based on sanger |
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|---|---|---|---|
| CN201610537027.3A CN107574236A (en) | 2016-07-02 | 2016-07-02 | It is a kind of that the method for assessing Alzheimer's disease risk is sequenced based on sanger |
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| CN201610537027.3A Pending CN107574236A (en) | 2016-07-02 | 2016-07-02 | It is a kind of that the method for assessing Alzheimer's disease risk is sequenced based on sanger |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111187825A (en) * | 2020-01-15 | 2020-05-22 | 丁玎 | Method and kit for detecting susceptibility sites of TOMM40 gene and APOE gene of Alzheimer's disease |
| CN111653360A (en) * | 2020-05-26 | 2020-09-11 | 苏州罗塞塔生物科技有限公司 | Intervention system based on brain health risk assessment |
-
2016
- 2016-07-02 CN CN201610537027.3A patent/CN107574236A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111187825A (en) * | 2020-01-15 | 2020-05-22 | 丁玎 | Method and kit for detecting susceptibility sites of TOMM40 gene and APOE gene of Alzheimer's disease |
| CN111653360A (en) * | 2020-05-26 | 2020-09-11 | 苏州罗塞塔生物科技有限公司 | Intervention system based on brain health risk assessment |
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| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180112 |