CN108103183B - Novel marker of the KCTD20 genes as early diagnosis parkinsonism - Google Patents
Novel marker of the KCTD20 genes as early diagnosis parkinsonism Download PDFInfo
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Abstract
The invention discloses parkinsonisms to diagnose related gene.The present invention screens the gene that expression is had differences in Parkinson's disease patients and normal person by genetic chip, proves that content of the KCTD20 genes in Parkinson's disease patients blood is higher than normal person by large sample QPCR verifications.It accordingly can be using KCTD20 genes as the biomarker of diagnosis parkinsonism.The achievement in research of the present invention provides a kind of noninvasive method clinically to carry out the early diagnosis of parkinsonism, is suitable for clinical expansion.
Description
Technical field
The invention belongs to molecular diagnosis fields, are related to a kind of molecular marker for parkinsonism diagnosis, and in particular to
Application of the molecular marker-KCTD20 genes in the product for preparing diagnosis parkinsonism in blood.
Background technology
Parkinson's disease (Parkinson Disease, PD) is also known as shaking plasy, is person in middle and old age's nervous system common disease.I
State's epidemiological survey, the illness rate of PD is the 1% (discussion of Luo Yi Primary ventricular hemorrhage drug therapies in 50 years old or more crowd
Liberation army physical-fitness medicine magazine, 2006,8 (1):1-5), men and women's morbidity no significant difference.Domestic and international statistical result is similar.With
The aging of society, it is estimated that the PD incidence to the year two thousand forty will increase by 4 times.The dyskinesia that PD is generated mainly has static
Property trembles, hypermyotonia, bradykinesia and abnormal posture etc..Often there is neuropsychological obstacle in end-stage disease；Or it is a series of
Motor complication.Late period disability is serious, can't take care of oneself, and heavy burden is brought to society and family.However cause PD really
It is still unclear so far to cut etiology and pathogenesis.It is now recognized that being by inherent cause, environmental factor joint effect, and at the age
Result (Schrag A, Ben-Shlomo Y, Quinn NP.Cross sectional prevalence under the effects that aging
survey of idiopathic Parkinson's disease and Parkinsonism in London[J]
.British Medical Journal.2000,321:21-2；Warner T,Schapira A H.Genetic and
environmental factors in the cause of Parkinson's disease[J].Annals of
Neurology.2003,53(13):16-23).Its major pathologic features are that substantia nigra of midbrain dopaminergic neuron largely loses
There is the inclusion body of referred to as Lewy body to be formed in the nerve cell for losing, and surviving.Lewy body includes the protein of many accumulations
And lipid, wherein α-syn (CooksonMR.The biochemistry of Parkinson's disease as the main component
[J].Annu Rev Biochem.2005,74:29-52).Clinically, diagnosis Parkinson's disease Main Basiss are the clinical conditions of patient
The experience of shape and doctor there is no the biochemical indicator for assisting in diagnosis Parkinson's disease.Currently, research hotspot both domestic and external is transferred to
On the biomarker for studying such degenerative disease, it is desirable to find stability and repeated good biomarker to help
Clinic early diagnosis.For disease, biomarker generally refer to for objective determination and evaluation a generic physiological,
Certain characteristic biochemical indicator in pathology or therapeutic process can know the life that body is presently in by the measurement to it
Process during object.More and more researchs find that specific gene can become the biological marker of diagnosis Parkinson
Object, such as the patent disclosure of following application number：201510713527.3,201510464905.9,201510463593.X,
201510463617.1,201510809837.5.Most of disease is all the disease of polygenes regulation and control, in order to improve examining for disease
Lots of genes Combining diagnosis is become inevitable by disconnected accuracy rate, it is therefore desirable to excavate more with the relevant molecular proportion mark of Parkinson's disease
Will object is so as to clinical application.
Invention content
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide one kind can be used for parkinsonism early diagnosis
Molecular marker.Compared to the diagnostic method of traditional parkinsonism, having for parkinsonism is diagnosed using gene marker
Promptness, specificity and sensitivity, for risk height, are taken to make patient that can know disease risks in disease early stage
Corresponding prevention and treatment measure.
To achieve the goals above, the present invention adopts the following technical scheme that：
The present invention provides application of the product of detection KCTD20 gene expressions in the tool for preparing diagnosis parkinsonism.
Further, the product of detection KCTD20 gene expressions mentioned above includes：Pass through RT-PCR, real-time quantitative
PCR, immune detection, in situ hybridization, chip or high-flux sequence detection of platform KCTD20 gene expression doses are to diagnose Parkinson
The product of disease.
Further, the product that parkinsonism is diagnosed with RT-PCR includes at least a pair of of specific amplified KCTD20 genes
Primer；The product that parkinsonism is diagnosed with real-time quantitative PCR includes at least the primer of a pair of of specific amplified KCTD20 genes；
It is described with immune detection diagnose parkinsonism product include：The antibody combined with KCTD20 protein-specifics；The original position
Hybridization diagnosis parkinsonism product include：With the probe of the nucleic acid array hybridizing of KCTD20 genes；It is described to diagnose pa with chip
The product of the gloomy disease of gold includes：Protein chip and genetic chip；Wherein, protein chip includes being combined with KCTD20 protein-specifics
Antibody, genetic chip include the probe with the nucleic acid array hybridizing of KCTD20 genes.
In specific embodiments of the present invention, the product that parkinsonism is diagnosed with real-time quantitative PCR includes at least
The sequence of the primer of a pair of of specific amplified KCTD20 genes is as shown in SEQ ID NO.1 and SEQ ID NO.2.
Preferably, the diagnostic tool includes chip, kit, test paper or high-flux sequence platform.Wherein, high pass measures
Sequence platform is a kind of special diagnostic tool, and the product of detection KCTD20 gene expressions can be applied to platform realization pair
The detection of the expression of KCTD20 genes.With the development of high throughput sequencing technologies, to the structure of the gene expression profile of a people
It builds to become and very easily work.By comparing the gene expression profile of Disease and normal population, it is easy which is analyzed
The exception of gene is related to disease.Therefore, know that the exception of KCTD20 genes is related to parkinsonism in high-flux sequence
Belong to the purposes of KCTD20 genes, equally within protection scope of the present invention.
The present invention also provides a kind of tools of diagnosis parkinsonism, and the diagnostic tool includes chip, kit, examination
Paper or high-flux sequence platform.
Wherein, the chip includes genetic chip, protein-chip；The genetic chip includes solid phase carrier and fixation
In the oligonucleotide probe of solid phase carrier, the oligonucleotide probe includes for detecting being directed to for KCTD20 gene transcription levels
The oligonucleotide probe of KCTD20 genes；The protein-chip includes solid phase carrier and is fixed on the KCTD20 of solid phase carrier
The specific antibody of albumen；The genetic chip can be used for detecting multiple genes including KCTD20 genes (for example, and pa
The gloomy relevant multiple genes of disease of gold) expression.The protein-chip can be used for detecting including KCTD20 albumen
The expression of multiple protein (such as with the relevant multiple protein of parkinsonism).By by multiple with parkinsonism mark
Will object detects simultaneously, is greatly improved the accuracy rate of parkinsonism diagnosis.
Wherein, the kit includes gene detecting kit and protein immunization detection kit；The genetic test examination
Agent box includes the reagent for detecting KCTD20 gene transcription levels；The protein immunization detection kit includes KCTD20 albumen
Specific antibody.Further, the reagent includes using RT-PCR, real-time quantitative PCR, immune detection, in situ hybridization or core
Piece method detects reagent needed for KCTD20 gene expression dose processes.Preference, the reagent include being directed to KCTD20 bases
The primer and/or probe of cause.It is easy to design according to the nucleotide sequence information of KCTD20 genes and can be used for detecting KCTD20
The primer and probe of gene expression dose.
Probe with the nucleic acid array hybridizing of KCTD20 genes can be DNA, RNA, DNA-RNA chimera, PNA or other
Derivative.There is no limit for the length of the probe, as long as completing specific hybrid, being specifically bound with purpose nucleotide sequence,
Any length can.The length of the probe can be as short as 25,20,15,13 or 10 bases longs.Equally, the probe
Length can be grown to 60,80,100,150,300 base-pairs or longer or even whole genes.Since different probe lengths is to miscellaneous
Efficiency, signal specificity is handed over to have different influences, the length of the probe is typically at least 14 base-pairs, and longest does not surpass generally
30 base-pairs are crossed, it is best with 15-25 base-pair with the length of purpose nucleotide sequence complementation.The probe self-complementary sequence
Row are most preferably less than 4 base-pairs, in order to avoid influence hybridization efficiency.
The high-flux sequence platform includes the reagent for detecting KCTD20 gene expression doses.
The test paper includes test paper carrier and the oligonucleotides that is fixed on test paper carrier, and the oligonucleotides can detect
The transcriptional level of KCTD20 genes.
Further, the specific antibody of the KCTD20 albumen includes monoclonal antibody, polyclonal antibody.The KCTD20
The specific antibody of albumen include complete antibody molecule, antibody any segment or modification (for example, chimeric antibody, scFv,
Fab, F (ab ') 2, Fv etc..As long as the segment can retain the binding ability with KCTD20 albumen.For protein water
When the preparation of flat antibody well known to a person skilled in the art, and the present invention may use any method it is described anti-to prepare
Body.
In specific embodiments of the present invention, the primer sequence for KCTD20 genes is as follows：Forward primer sequence
Row are as shown in SEQ ID NO.1, and reverse primer is as shown in SEQ ID NO.2.
The source of KCTD20 genes and its expression product for diagnosing parkinsonism includes but not limited to blood, tissue
Liquid, urine, saliva, spinal fluid etc. can obtain the body fluid of genomic DNA.In specific embodiments of the present invention, for examining
The KCTD20 genes of disconnected parkinsonism and its source of expression product are blood.
In the context of the present invention, " KCTD20 genes " includes any function of KCTD20 genes and KCTD20 genes
The polynucleotides of equivalent.KCTD20 genes (Chromosome 6, NC_000006.12 (36442767..36491143)) sequence
It is inquired in the international public GenBank GeneBank of Lie Ke.
In the context of the present invention, KCTD20 gene expression products include the portion of KCTD20 albumen and KCTD20 albumen
Divide peptide.The partial peptide of the KCTD20 albumen contains and the relevant functional domain of parkinsonism.
" KCTD20 albumen " includes any functional equivalent of KCTD20 albumen and KCTD20 albumen.The function is equivalent
Object includes KCTD20 albumen conservative variation protein or its active fragment or its reactive derivative, allelic variant, natural
Mutant, induced mutants, can be with the encoded protein of DNA of the DNA hybridization of KCTD20 under high or low stringent condition.
It is known that, conventionally, the modification of one or more amino acid does not interfere with the function of protein in a protein.
Those skilled in the art can approve the amino acid for changing single amino acids or small percentage or to amino acid sequence it is individual add,
Missing, insertion, replacement are conservative modifications, and the change of wherein protein generates the protein with identity function.Function phase is provided
As the Conservative substitution tables of amino acid be well known in the art.
Example by adding the protein of an amino acid or more amino acid modification is melting for KCTD20 albumen
Hop protein.For the peptide or protein with KCTD20 protein fusions, there is no limit as long as the fusion protein of gained retains
The biological activity of KCTD20 albumen.
In the context of the present invention, " diagnosis parkinsonism " had both included judging whether subject has suffered from Parkinson
Disease also includes the risk that judges subject and whether there is with parkinsonism.
The advantages of the present invention：
Present invention firstly discovers that KCTD20 gene expressions are related to parkinsonism, pass through and detect KCTD20 in subject
Expression, it can be determined that whether subject suffers from parkinsonism or judges that subject whether there is the risk with parkinsonism,
To instruct clinician to provide prevention scheme or therapeutic scheme to subject.
Present invention finds a kind of new molecular marked compound-KCTD20 genes, compare traditional detection means, gene diagnosis
More in time, more special, sensitiveer, the early diagnosis of parkinsonism can be realized, to reduce the death rate of parkinsonism.
Description of the drawings
Fig. 1, which is shown, utilizes differential expression of the genechip detection KCTD20 genes in Parkinson's disease patients and normal person；
Fig. 2 shows the differential expression in Parkinson's disease patients and normal person using QPCR detection KCTD20 genes.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this
It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip
Part, such as Sambrook et al., molecular cloning：Laboratory manual (New York:Cold Spring HarborLaboratory
Press, 1989) condition described in, or according to the normal condition proposed by manufacturer.
Embodiment 1 screens the gene of differential expression in Parkinson's disease patients and normal person
1, research object：
Collect primary PD patient 10, man 5, female 5, age 32-85 Sui, the course of disease -20 years 5 months.PD enters a group mark
It is accurate：Diagnostic criteria meets PD clinical criterias (with reference to " Jiang Yuping, Wang Jian, Ding Zhengtong, etc. Primary ventricular hemorrhage is examined
Disconnected standard, 2005, Chinese Clinical Neuscience, 2006,14:40").Exclusion criteria：(1) essential tremor；(2) secondary pa
The gloomy syndrome of gold；(3) advanced dementia, dysarthrosis person；(4) other mental disease persons are suffered from.This studies oneself and passes through hospital's human relations
The reason committee ratifies and all patients sign informed consent form.
Normal group：Choose the healthy volunteer 10 at age 32-80 Sui, each 5 of men and women.
Age, the not statistically significant (P of gender differences between two groups>0.10) it, is comparable.
2, in blood total serum IgE extraction
(1) homogenized (Homogenization)
Fresh blood (peripheral blood) is directly taken, 3 times of volume erythrocyte cracked liquids are added, 10 points are placed at room temperature for after mixing
Clock, 10,000rpm centrifugations 1 minute.It thoroughly inhales and abandons supernatant, collect leukocyte cell pellet.Per the leucocyte of 100-200 μ l blood collections
1ml TRIzol are added in precipitation.
(2) it is layered (Phase Separation)
A. after TRIzol is added in sample, it is placed at room temperature for 5min, sample is made fully to crack.
B. 200 μ l chloroforms are added per 1ml TRIzol, 3-5min is placed at room temperature for after shaking vigorously and mix well makes its natural split-phase.
(3) RNA precipitate (RNAPrecipitation)
A.4 DEG C 12,000rpm centrifuges 10-15min.Sample can be divided into three layers：The organic phase of yellow, middle layer and colourless
Water phase, RNA mainly in water phase, water phase (can usually draw 550 μ l) are transferred in new pipe.
B. isometric ice-cold isopropanol is added in supernatant, is placed at room temperature for 10-20min.4 DEG C of 12,000rpm centrifugations
10min abandons supernatant, and RNA precipitate is in tube bottom.
(4) RNA rinses (RNA Wash)
75% ethyl alcohol of 1ml (being prepared with RNase-free water) is added in a.RNA precipitations, mildly vibrates centrifuge tube, it is heavy to suspend
It forms sediment.75% ethyl alcohol of 1ml is added per 1ml TRIzol.
B.4 DEG C 5,000-8,000rpm centrifuges 1-2min, abandons supernatant；Of short duration rapid centrifugation is carefully inhaled with pipettor and is abandoned
Clearly, 1-2 minutes are placed at room temperature for and dries precipitation.
(5) dissolving RNA (Redissolving the RNA)
50-100 μ l RNase-free water is added in precipitation, flicks tube wall, fully to dissolve RNA, -70 DEG C of preservations.
3, RNA mass and purity detecting
RNA mass：It is indicated by RNA integralities, plain agar sugar gel electrophoresis (deposition condition can be used：1.2% glue；
0.5 × TBE electrophoretic buffers；150v, 15 minutes) detection integrality.
RNA purity：OD260/OD280 ratios are the indexs for weighing protein contamination degree in RNA sample.High quality
RNA sample, OD260/OD280 values (10mM Tris, pH7.5) are 2.0 or so.
4, gene chip hybridization and scanning
After the linearized amplification of total serum IgE, cy3-UTP is marked, and the cRNAs after fluorescent marker uses RNEASY Mini Kit
Purifying, fragmentation processing is carried out with the RNA Fragmentation Reagents of Amhion to the cRNAs marked.Using U.S.
People's full genome chip of expression spectrum (4x 44K genes) of Agilent companies of state, 65 DEG C of hybridization 17h in chip hybridization stove, then
Elution, dyeing, finally use Agilent DNA MicroarrayScanner scanner scannings.
5, chip data processing and analysis
Chip after hybridization imports data to analysis software, for two groups of ratios after chip scanner reads data point
Natural logrithm absolute value be more than 2.0 or the gene less than 0.5 as difference expression gene.
6, statistical procedures
Data analysis is carried out using 13.0 statistical softwares of SPSS, group difference compares using one-way analysis of variance method, P<
The significant meaning of 0.05 difference.
7, result
As a result (as shown in Figure 1) is shown, compared with normal person, the mRNA water of KCTD20 genes in Parkinson's disease patients blood
HUD, which writes, to be increased, and difference has statistical significance (P<0.05).
The gene of differential expression in 2 QPCR experimental verifications Parkinson's disease patients of embodiment and normal person
1, research object：
Screening criteria is the same as embodiment 1, Parkinson's disease patients and each 50 of normal person.
2, in blood total serum IgE extraction
Step is the same as embodiment 1.
3, reverse transcription
Reverse transcription is carried out to l μ g total serum IgEs with RT Buffer and synthesizes cDNA.Using 25 μ l reaction systems, each sample
It takes 1 μ g total serum IgEs as template ribonucleic acid, following components is separately added into PCR pipe：DEPC water, 5 × RT Buffer,
10mmol/L dNTP, 0.1mmol/l DTT, 30 μm of mol/l Oligo dT, 200U/ μ l M-MLV, template ribonucleic acid.42 DEG C of incubations
1h, 72 DEG C of 10min, of short duration centrifugation.
4、QPCR
(1) design of primers
QPCR amplimers are designed according to the coded sequence of KCTD20 genes and GAPDH genes in Genbank, are given birth to by Shanghai
Work biotechnology Services Co., Ltd synthesizes.Specific primer sequence is as follows：
KCTD20 genes：
Forward primer is 5 '-GCTGCTTAGTGGATGATAC-3 ' (SEQ ID NO.1)；
Reverse primer is 5 '-AGATTATGAGGACCAGATGA-3 ' (SEQ ID NO.2),
GAPDH genes：
Forward primer is 5 '-TTTAACTCTGGTAAAGTGGATAT-3 ' (SEQ ID NO.3)；
Reverse primer is 5 '-GGTGGAATCATATTGGAACA-3 ' (SEQ ID NO.4).
(2) PCR reaction systems are prepared according to table 1：
Wherein, SYBR Green PCRs system is purchased from Invitrogen companies.
1 PCR reaction systems of table
(3) PCR reaction conditions：95 DEG C of 10min, (95 DEG C of 10s, 60 DEG C of 40s) * 45 cycles.Using SYBR Green as
Fluorescent marker carries out PCR reactions on Light Cycler fluorescence quantitative PCR instruments, true by melt curve analysis analysis and electrophoresis
Determine purpose band, Δ Δ CT methods carry out relative quantification.
5, statistical method
Result data is indicated in a manner of mean+SD, is united using SPSS13.0 statistical softwares
Meter analysis, difference between the two is examined using t, it is believed that works as P<There is statistical significance when 0.05.
6, result
The results are shown in Figure 2, and compared with normal person, the mRNA level in-site of KCTD20 genes is notable in Parkinson's disease patients blood
Increase, difference has statistical significance (P<0.05), as a result homogenic array experiment.
3 immunoblot experiment of embodiment verifies the expression product of difference expression gene in Parkinson's disease patients and normal person
1, clinical subjects：With embodiment 2.
2, monocyte detaches
Parkinson's disease patients and normal person extracting vein blood 10ml, injection are contained in the sterile vials of heparin, light immediately after capping
Jog is even.Isometric HBSS (NaCl 8.0g, Na is added with aseptic straw2HPO40.132g, KH2PO40.06g, KCl
0.4g, phenol red 1ml, NaHCO30.35g, D-Glucose 1.0g are dissolved in 1000ml distilled waters), to reduce the cohesion of red blood cell.
It draws 8ml lymphocytes separating solutions to set in 50ml centrifuge tubes, dilute blood is slowly added to along tube wall, keep interface to understand, not
So that the two is mutually mixed, 30min, the careful canescence for drawing layering liquid and blood plasma handing-over position muddiness are centrifuged in 20 DEG C of 2000r/min
Layer, i.e. buffy coat are added in another centrifuge tube, and 2 times are washed with the HBSS of 5 times of volumes, successively with 2000r/min,
1500r/min centrifuges 10min at room temperature, the blood platelet largely mixed to remove, with 10ml distilled waters and cell mass
1min is mixed, residual red blood cells is made to crack, is then rapidly added equivalent 1.8%NaCl solution, supernatant is removed in 2000r/min centrifugations,
Cell is adjusted to 1 × 10 with HBSS solution after cell count6A/ml is spare.
3, monocyte gross protein extracts
By cell suspension (a concentration of 1 × 10 obtained by above-mentioned experiment6A/ml) room temperature 1 000r/min centrifuge 10min, abandon
It is added 100 μ l lysis buffers after clear, 4 DEG C of concussion 1h, with ultrasonoscope smudge cells, each 10s, totally 10 times, in 4 DEG C
12000r/min centrifuges 1h；Supernatant Brandford standard measure albumen is taken, is distributed into 2.5 μ g/ μ l, -80 DEG C of refrigerators preserve standby
With.
4, Western blot are detected
Total protein of cell Brandford standard measures take to mix with sample buffer in right amount and boil 5min, cooling 5min；
It takes 30pg albumen to be loaded to 15% polyacrylamide gel prepared, carries out electrophoresis, start to be set as 80V constant pressures, see
120V is increased to after Marker；Glue after electrophoresis is taken out, 50min is shifted in 100V using the half-dried transferring systems of Bio.Rad；Turn
It after film, is washed once with 1xPBS, immerses confining liquid, 40C is overnight；Confining liquid is outwelled, Western cleaning solutions are added and wash 5-
10min is added primary antibody shaking table room temperature and hybridizes 2h；According to proper proportion Block buffer is diluted in Western secondary antibody diluents
In, it is incubated 60min；Film washing liquid is washed 3 times, each 10min；Use the development of ECL reagents, fixing detection protein expression.
5, statistical procedures
The gray value of protein band is analyzed using Image J softwares, using β-actin as internal reference, by purpose informal voucher
The gray value of band is normalized.Result data is indicated in a manner of mean+SD, is used
SPSS13.0 statistical softwares are next for statistical analysis, and difference between the two is examined using t, it is believed that works as P<There is system when 0.05
Meter learns meaning.
6, result
The results show that with the albumen relative expression levels in normal person's group blood for 1, KCTD20 in parkinsonism group blood
Protein level is 5.87 ± 1.45, is significantly increased, and difference has statistical significance (P<0.05).
The explanation of above-described embodiment is only intended to understand the method and its core concept of the present invention.It should be pointed out that for this
For the those of ordinary skill in field, without departing from the principle of the present invention, several improvement can also be carried out to the present invention
And modification, these improvement and modification will also be fallen into the protection domain of the claims in the present invention.
Sequence table
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Claims (10)
1. detecting application of the product of KCTD20 gene expressions in the tool for preparing diagnosis parkinsonism.
2. application according to claim 1, which is characterized in that the product includes：By RT-PCR, real-time quantitative PCR,
Immune detection, in situ hybridization, chip or high-flux sequence detection of platform KCTD20 gene expression doses are to diagnose parkinsonism
Product.
3. application according to claim 2, which is characterized in that the product for diagnosing parkinsonism with RT-PCR at least wraps
Include the primer of a pair of of specific amplified KCTD20 genes；The product that parkinsonism is diagnosed with real-time quantitative PCR includes at least one
To the primer of specific amplified KCTD20 genes；It is described with immune detection diagnose parkinsonism product include：With KCTD20 albumen
The antibody of specific binding；It is described in situ hybridization diagnose parkinsonism product include：With the nucleic acid sequence of KCTD20 genes
The probe of hybridization；It is described with chip diagnose parkinsonism product include：Protein chip and genetic chip；Wherein, protein chip
Include the antibody combined with KCTD20 protein-specifics, genetic chip includes the spy with the nucleic acid array hybridizing of KCTD20 genes
Needle.
4. application according to claim 3, which is characterized in that the product for diagnosing parkinsonism with real-time quantitative PCR
The primer of a pair of of the specific amplified KCTD20 genes included at least is as shown in SEQ ID NO.1 and SEQ ID NO.2.
5. application according to claim 1, which is characterized in that the tool can be by detecting KCTD20 genes in sample
Expression diagnose parkinsonism.
6. application according to claim 5, which is characterized in that the tool includes chip, kit, test paper or high throughput
Microarray dataset.
7. application according to claim 6, which is characterized in that the chip includes genetic chip, protein-chip；It is described
Genetic chip includes solid phase carrier and is fixed on the oligonucleotide probe of solid phase carrier, and the oligonucleotide probe includes being used for
Detect the oligonucleotide probe for KCTD20 genes of KCTD20 gene transcription levels；The protein-chip includes that solid phase carries
Body and be fixed on solid phase carrier KCTD20 albumen specific antibody；The kit includes gene detecting kit and egg
White immunity detection reagent；The gene detecting kit includes the reagent for detecting KCTD20 gene transcription levels；It is described
Protein immunization detection kit includes the specific antibody of KCTD20 albumen；The test paper includes turning for detecting KCTD20 genes
Record horizontal reagent；The high-flux sequence platform includes the reagent for detecting KCTD20 gene transcription levels.
8. application according to claim 7, which is characterized in that it is described detection KCTD20 gene transcription levels reagent include
For the primer and/or probe of KCTD20 genes.
9. application according to claim 8, spy are characterized in that, the primer sequence for KCTD20 genes is as follows：
Forward primer sequence is as shown in SEQ ID NO.1, and reverse primer is as shown in SEQ ID NO.2.
10. according to the application described in any one of claim 5-9, which is characterized in that the sample is blood.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711481574.5A CN108103183B (en) | 2017-12-29 | 2017-12-29 | Novel marker of the KCTD20 genes as early diagnosis parkinsonism |
| PCT/CN2018/123794 WO2019129029A1 (en) | 2017-12-29 | 2018-12-26 | Gene as marker for diagnosing and treating parkinson's disease and use thereof |
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| CN201711481574.5A CN108103183B (en) | 2017-12-29 | 2017-12-29 | Novel marker of the KCTD20 genes as early diagnosis parkinsonism |
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Denomination of invention: KCTD20 gene serving as new type marker for early diagnosing Parkinsonism Effective date of registration: 20181225 Granted publication date: 20181102 Pledgee: Huaxia Bank Beijing branch Wanliu Limited by Share Ltd Pledgor: Beijing Yang Shen biology information technology company limited Registration number: 2018990001252 |
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Effective date of registration: 20191227 Address after: Room 2503, Qianshan building, building D2, Qingdao International Innovation Park Phase II, No. 1, Keyuan Weiyi Road, Laoshan District, Qingdao, Shandong Province Patentee after: Qingdao Yangshen biomedical Co., Ltd Address before: 100080 Beijing city Haidian District Shanyuan Street No. 1 cubic court building room 3103 Patentee before: Beijing Yang Shen biology information technology company limited |