CN107988347A - A kind of detection method of alzheimer's disease tumor susceptibility gene and its application - Google Patents
A kind of detection method of alzheimer's disease tumor susceptibility gene and its application Download PDFInfo
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- CN107988347A CN107988347A CN201711369901.8A CN201711369901A CN107988347A CN 107988347 A CN107988347 A CN 107988347A CN 201711369901 A CN201711369901 A CN 201711369901A CN 107988347 A CN107988347 A CN 107988347A
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Abstract
To comprise the following steps the invention discloses the present invention provides the detection method of a kind of detection method of alzheimer's disease tumor susceptibility gene and its application, the alzheimer's disease tumor susceptibility gene：Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded, obtain amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；Step 2: carrying out unimolecule amplification to sequencing library, multiple unimolecule amplified productions corresponding with the multiple target area are obtained；Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the sequence information of the multiple target area is obtained.The multiple regions of alzheimer's disease tumor susceptibility gene can be carried out at the same time detection, greatly reduce testing cost by a kind of detection method of alzheimer's disease tumor susceptibility gene provided by the invention.
Description
Technical field
The invention belongs to genetic engineering field, and in particular to a kind of detection method of alzheimer's disease tumor susceptibility gene and its
Using.
Background technology
Tumor susceptibility gene refers to there is certain associated gene or allele with specified disease.To the susceptible of complex disease
Gene and mutational site are detected, and can obtain its specific gene sequence information.On this basis, can combine subsequently into
Molecular biology test, clinical test, clinical observation and the statistical analysis of integrated data of one step, establish disease model, so that
Realize the neurological susceptibility of earlier evaluations disease and take the occurrence probability of corresponding precautionary measures reduction disease.Numerous studies confirm to lose
Biography factor plays a significant role in the neurological susceptibility of disease, the inherent cause of complex disease oneself be confirmed the existence of minor gene dosage
The cumulative of effect, i.e. minor gene effect can just form obvious phenotype comprehensive effect.Therefore, to the multiple susceptible of disease
Gene and mutational site be detected it is most important, especially for most important genetic mutation form in genome --- it is single
Nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNP), it accounts for genetic mutation in whole gene group
92%.
Alzheimer's disease is called senile dementia, is a kind of central nervous system degeneration, insidious onset, the course of disease is in slow
Property progressive, is senile dementia the most common type type.Be mainly shown as gradual memory obstacle, cognition dysfunction,
The neuropsychic symptom such as personality change and aphasis, seriously affects social, occupation and vital function.The disease of alzheimer's disease
Cause and pathogenesis not yet illustrate, and this area lacks comprehensive, system, effective recognition methods, for grinding for alzheimer's disease
It is also less to study carefully achievement.
The content of the invention
In view of the deficiencies of the prior art, it is a primary object of the present invention to provide a kind of alzheimer's disease tumor susceptibility gene
Detection method and its application.
To realize aforementioned invention purpose, the technical solution adopted by the present invention includes：The present invention provides a kind of A Erzi seas
The detection method of silent disease tumor susceptibility gene, it comprises the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
Among some embodiments, the gene order of the target area is SEQ ID NO：1 and SEQ ID NO：2.
Among some embodiments, the tumor susceptibility gene is MTHFR.
Among some embodiments, the step 1 includes：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component is using any one in protruding terminus connector, connector and Y connection with loop-stem structure or two or more
Combination.
Among some more specific embodiment, the step B comprises the following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
Present invention also offers a kind of kit gene, which examines in above-mentioned alzheimer's disease tumor susceptibility gene
Application in survey method.
Among some embodiments, the kit gene include be used for multiple target areas in sample to be tested into
The alzheimer's disease tumor susceptibility gene specific primer of row amplification；And
Joint component, for being combined structure sequencing library with amplified production.
Among some more specific embodiment, the joint component is using protruding terminus connector, band loop-stem structure
Connector and at least one of Y connection.
Compared with prior art, advantages of the present invention includes：A kind of alzheimer's disease tumor susceptibility gene provided by the invention
Detection method, detection can be carried out at the same time to the multiple regions of alzheimer's disease tumor susceptibility gene, greatly reduce testing cost；
In addition, the sample for including heterozygote can be identified in the alzheimer's disease tumor susceptibility gene detection method of the present invention, carry
High detection sensitivity.
Embodiment
More detailed explanation will hereafter be made to technical scheme.It is understood, however, that in model of the present invention
In enclosing, above-mentioned each technical characteristic of the invention and below（Such as embodiment）Can between each technical characteristic of middle specific descriptions
It is combined with each other, so as to form new or preferable technical solution.As space is limited, not repeated them here.
The present invention provides a kind of detection method of alzheimer's disease tumor susceptibility gene, it comprises the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
Among some embodiments, the gene order of the target area is SEQ ID NO：1 and SEQ ID NO：2.
Among some embodiments, the tumor susceptibility gene is MTHFR.
Among some embodiments, the step 1 includes：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component is using any one in protruding terminus connector, connector and Y connection with loop-stem structure or two or more
Combination.
Among some more specific embodiment, the step B comprises the following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
Present invention also offers a kind of kit gene, which examines in above-mentioned alzheimer's disease tumor susceptibility gene
Application in survey method.
Among some embodiments, the kit gene include be used for multiple target areas in sample to be tested into
The alzheimer's disease tumor susceptibility gene specific primer of row amplification；And
Joint component, for being combined structure sequencing library with amplified production.
Among some more specific embodiment, the joint component is using protruding terminus connector, band loop-stem structure
Connector and at least one of Y connection.
Embodiment 1
A kind of detection method of alzheimer's disease tumor susceptibility gene, it comprises the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
Among some embodiments, the gene order of the target area is SEQ ID NO：1 and SEQ ID NO：2.
Among some embodiments, the tumor susceptibility gene is MTHFR.
Among some embodiments, the step 1 includes：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component uses protruding terminus connector.
Among some more specific embodiment, the step B comprises the following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
Present invention also offers a kind of kit gene, which examines in above-mentioned alzheimer's disease tumor susceptibility gene
Application in survey method.
Among some embodiments, the kit gene include be used for multiple target areas in sample to be tested into
The alzheimer's disease tumor susceptibility gene specific primer of row amplification；And
Joint component, for being combined structure sequencing library with amplified production.
Among some more specific embodiment, the joint component uses protruding terminus connector.
Embodiment 2
A kind of detection method of alzheimer's disease tumor susceptibility gene, it comprises the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
Among some embodiments, the gene order of the target area is SEQ ID NO：1 and SEQ ID NO：2.
Among some embodiments, the tumor susceptibility gene is MTHFR.
Among some embodiments, the step 1 includes：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component uses the connector with loop-stem structure.
Among some more specific embodiment, the step B comprises the following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
Present invention also offers a kind of kit gene, which examines in above-mentioned alzheimer's disease tumor susceptibility gene
Application in survey method.
Among some embodiments, the kit gene include be used for multiple target areas in sample to be tested into
The alzheimer's disease tumor susceptibility gene specific primer of row amplification；And
Joint component, for being combined structure sequencing library with amplified production.
Among some more specific embodiment, the joint component uses the connector with loop-stem structure.
Embodiment 3
A kind of detection method of alzheimer's disease tumor susceptibility gene, it comprises the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
Among some embodiments, the gene order of the target area is SEQ ID NO：1 and SEQ ID NO：2.
Among some embodiments, the tumor susceptibility gene is MTHFR.
Among some embodiments, the step 1 includes：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component uses Y connection.
Among some more specific embodiment, the step B comprises the following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
Present invention also offers a kind of kit gene, which examines in above-mentioned alzheimer's disease tumor susceptibility gene
Application in survey method.
Among some embodiments, the kit gene include be used for multiple target areas in sample to be tested into
The alzheimer's disease tumor susceptibility gene specific primer of row amplification；And
Joint component, for being combined structure sequencing library with amplified production.
Among some more specific embodiment, the joint component uses Y connection.
The technology contents and technical characteristic of the present invention have revealed that as above, but those skilled in the art still may base
Make a variety of replacements and modification without departing substantially from spirit of the present invention, therefore, the scope of the present invention in teachings of the present invention and announcement
The revealed content of embodiment should be not limited to, and various replacements and modification without departing substantially from the present invention should be included, and is this patent Shen
Please claim covered.
<110>The vast health consultation management of light（Shanghai）Co., Ltd
<120>A kind of detection method of alzheimer's disease tumor susceptibility gene and its application
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 246
<212> DNA
<213> Artificial
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<223>Artificial sequence
<400> 1
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tgcctggtga tcagagttgt ttttcattct cttttcttct tttagccttg cctccccgat 180
tgaaagagat gaaaagccag gaatcggctg caggttccaa actagtcctt cggtgtgaaa 240
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<213> Artificial
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tttcatattg aatttatatt tagcctgttc tagtacatag tctctgcata gtatatccat 180
ggcatcc 187
Claims (8)
1. a kind of detection method of alzheimer's disease tumor susceptibility gene, it is characterised in that comprise the following steps：
Step 1: using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are carried out
Amplification, obtains amplified production corresponding with the multiple target area, and based on amplified production structure sequencing library；
Step 2: carrying out unimolecule amplification to sequencing library, obtain multiple unimolecules corresponding with the multiple target area and expand
Increase production thing；
Step 3: carrying out high throughput gene sequencing to the multiple unimolecule amplified production at the same time, the multiple target area is obtained
The sequence information in domain.
2. the detection method of alzheimer's disease tumor susceptibility gene according to claim 1, it is characterised in that：The target area
Gene order be SEQ ID NO：1 and SEQ ID NO：2.
3. the detection method of alzheimer's disease tumor susceptibility gene according to claim 1, it is characterised in that：The tumor susceptibility gene
For MTHFR.
4. the detection method of alzheimer's disease tumor susceptibility gene according to claim 1, it is characterised in that the step 1 bag
Include：
Step A, using alzheimer's disease tumor susceptibility gene specific primer, multiple target areas in sample to be tested are expanded
Increase, obtain amplified production corresponding with the multiple target area；
Step B, it is attached using joint component, amplified production corresponding with the multiple target area, obtains sequencing library；
The joint component is using any one in protruding terminus connector, connector and Y connection with loop-stem structure or two or more
Combination.
5. the detection method of alzheimer's disease tumor susceptibility gene according to claim 4, it is characterised in that the step B bags
Include following steps：
1., pair corresponding with a multiple target area amplified production carry out fragmentation, obtain fragmentation products；
2., using joint component, be attached with the fragmentation products, build sequencing library.
A kind of 6. kit gene, it is characterised in that：The kit gene is in A Erzi seas any one of claim 1-5
Application in silent disease tumor susceptibility gene detection method.
7. kit gene according to claim 6, it is characterised in that the kit gene includes being used to treat test sample
The alzheimer's disease tumor susceptibility gene specific primer that multiple target areas in product are expanded；And joint component, it is used for
Structure sequencing library is combined with amplified production.
8. kit gene according to claim 7, it is characterised in that the joint component using protruding terminus connector,
At least one of connector and Y connection with loop-stem structure.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109215738A (en) * | 2018-10-12 | 2019-01-15 | 中南大学 | The prediction technique of Alzheimer's disease related gene |
| CN111593107A (en) * | 2020-01-15 | 2020-08-28 | 丁玎 | Detection method and kit for susceptibility sites rs10119 and rs71352238 of Alzheimer's disease |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1548552A (en) * | 2003-05-20 | 2004-11-24 | 岱 张 | Detection method and use of susceptible gene of schizophrenia |
| CN102373288A (en) * | 2011-11-30 | 2012-03-14 | 盛司潼 | Method and kit for sequencing target areas |
-
2017
- 2017-12-19 CN CN201711369901.8A patent/CN107988347A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1548552A (en) * | 2003-05-20 | 2004-11-24 | 岱 张 | Detection method and use of susceptible gene of schizophrenia |
| CN102373288A (en) * | 2011-11-30 | 2012-03-14 | 盛司潼 | Method and kit for sequencing target areas |
Non-Patent Citations (1)
| Title |
|---|
| LEILA MANSOURI等: "Association of methylenetetrahydrofolate reductase polymorphisms with susceptibility to Alzheimer"s disease", 《CLINICAL NEUROLOGY AND NEUROSURGERY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109215738A (en) * | 2018-10-12 | 2019-01-15 | 中南大学 | The prediction technique of Alzheimer's disease related gene |
| CN109215738B (en) * | 2018-10-12 | 2020-04-28 | 中南大学 | Method for predicting Alzheimer's disease-related gene |
| CN111593107A (en) * | 2020-01-15 | 2020-08-28 | 丁玎 | Detection method and kit for susceptibility sites rs10119 and rs71352238 of Alzheimer's disease |
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