CN106636408A - Budd-Chiari syndrome diagnosis tool - Google Patents
Budd-Chiari syndrome diagnosis tool Download PDFInfo
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- CN106636408A CN106636408A CN201611222906.3A CN201611222906A CN106636408A CN 106636408 A CN106636408 A CN 106636408A CN 201611222906 A CN201611222906 A CN 201611222906A CN 106636408 A CN106636408 A CN 106636408A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/08—Hepato-biliairy disorders other than hepatitis
- G01N2800/085—Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/50—Determining the risk of developing a disease
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/54—Determining the risk of relapse
Abstract
The invention belongs to the field of gene diagnosis, and particularly relates to an application of a GFI1B gene to preparation of a tool for diagnosing a Budd-Chiari syndrome. The GFI1B gene can be taken as a specific marker gene for diagnosing the Budd-Chiari syndrome, so that more accurate and rapid diagnosis of the Budd-Chiari syndrome is realized.
Description
Technical field
The present invention relates to medical diagnosis on disease field, more particularly it relates to a kind of Bgudd-Chiari Syndrome diagnostic tool.
Background technology
Budd-Chiari syndrome (Budd-Chiari syndrome, BCS) is with epimere cavity of resorption by vena hepatica and (or) its opening
What vein occlusion pathology caused is often accompanied by property portal hypertension after a kind of liver with the characteristics of inferior vena cava syndrome.From global model
From the point of view of enclosing, the BCS incidences of disease are relatively low, and its cause of disease has obvious regional difference, and western countries are common with hepatic venous obstruction type BCS, greatly
There are the clear and definite cause of disease, such as oral contraceptive, gestation, blood disease more, have document report, vena hepatica type BCS and gene
High blood coagulation state, Hepatic venous thrombosis are relevant caused by mutation；And in Asia and Southern African region then with IVC obstructive type BCS
Common, pathogenic factor is mostly unclear.
BCS can cause liver damage, portal hypertension, even result in cirrhosis, hepatic failure, UGB etc. serious
Complication, Nature prognosis extreme difference, survival rate is very low within 5 years, and the non-operative treatment death rate is high.The clinical manifestation of BCS patient's early stage is hidden
Hide, without specificity, and lack the effective ways of early stage diagnosis and treatment, patient when medical more belong to late period, complicated clinical manifestation is various, holds
Easily cause mistaken diagnosis, control by mistake.The domestic research to BCS focuses mostly in terms of clinical treatment, by contrast epidemiology and teiology
Research it is delayed, lack the whole nation generaI investigation and Study of Etiology.Therefore, carry out BCS Study of Etiology in a deep going way, improve the early stage of BCS
Diagnosis is the key issue for improving BCS survival of patients and prognosis.
The content of the invention
The present invention relates to a kind of diagnostic tool of Bgudd-Chiari Syndrome, the diagnostic tool is by diagnosing molecular marker
The expression of GFI1B is playing diagnostic function.The present invention is experimentally confirmed the GFI1B bases in the blood of Bgudd-Chiari Syndrome patient
Because mRNA contents are significantly higher than normal population, therefore the property of the differential expression of GFI1B genes becomes and can be used to diagnose
Whether experimenter suffers from the molecular marker of Bgudd-Chiari Syndrome.
Specifically, the invention provides application of the product of detection GFI1B in Bgudd-Chiari Syndrome diagnostic tool is prepared.
Further, the product of the detection GFI1B includes the product of GFI1B gene expression amounts in detection sample.
Further, the product of the detection GFI1B include can quantitatively in sample GFI1B gene mRNAs product, and/
Or can quantitatively in sample GFI1B albumen product.
The product of GFI1B gene mRNAs can be based on using the known method of nucleic acid molecules to send out in the quantitative sample of the present invention
Wave its function：Such as PCR, such as Southern hybridization, Northern hybridization, dot blot, FISH (FISH), the micro- battle arrays of DNA
Row, ASO methods, high-flux sequence platform etc..Can qualitatively, quantitatively or semi-quantitatively implement analysis using the product.
The nucleic acid being included in the said goods can be obtained by chemical synthesis, or be contained by preparing from biomaterial
Expect the gene of nucleic acid, then expand it to obtain using the primer for being designed for expanding expectation nucleic acid.
Further, the PCR method is known method, for example, ARMS (Amplification Refractory
Abruptly-changing system is not answered in Mutation System, amplification) method, RT-PCR (reverse transcriptase-PCR) method, nesting PCR methods etc..Amplification
Nucleic acid can be by using dot blotting hybridization method, Surface Plasmon Resonance (SPR methods), PCR-RFLP methods, original position RT-PCR
Method, PCR-SSO (sequence specific oligonucleotide) method, PCR-SSP methods, AMPFLP (amplifiable fragment length polymorphism) method,
MVR-PCR methods and PCR-SSCP (single-strand conformation polymorphism) methods are detecting.
It is described can quantitatively the product of GFI1B gene mRNAs includes the specific amplified used in real-time quantitative PCR in sample
The primer of GFI1B genes, the primer sequence is as shown in SEQ ID NO.1 and SEQ ID NO.2.
The primer that product includes can be prepared by chemical synthesis, by using those skilled in the art will know that side
Method is suitably designed with reference to Given information, and is prepared by chemical synthesis.
Nucleic acid recited above may also include probe, and the probe can be prepared by chemical synthesis, by using this
The method that art personnel know appropriately is designed with reference to Given information, and is prepared by chemical synthesis, or can be led to
Cross from biomaterial and prepare containing the gene for expecting nucleotide sequence, and expanded using the primer for being designed for expanding expectation nucleotide sequence
Increase it to prepare.
The product of GFI1B albumen can be based on and its work(is played using the known method of antibody in the quantitative sample of the present invention
Energy：For example, ELISA, radioimmunoassay, immunohistochemical method, western blot etc. can be included.
The product of GFI1B albumen includes the antibody or its piece of specific binding GFI1B albumen in the quantitative sample of the present invention
Section.The antibody or its fragment of any structure, size, immunoglobulin class, origin etc. can be used, as long as it combines target protein
Matter.The antibody or its fragment that the detection product of the present invention includes can be monoclonal or polyclonal.Antibody fragment
Refer to and retain peptide of the antibody to an antibody part (Partial Fragment) of the binding activity of antigen or containing an antibody part.Antibody fragment
F (ab ') can be included2, Fab ', Fab, scFv (scFv), disulphide bonding Fv (dsFv) or its polymer, dimerization
V areas (double antibody) or the peptide containing CDR.The present invention quantitative sample in GFI1B albumen product can include encoding antibody or
The detached nucleic acid of the amino acid sequence of Encoding Antibody Fragment, the carrier comprising the nucleic acid, and carry the cell of the carrier.
Antibody can be by well known to a person skilled in the art method be obtaining.For example, prepare and retain target all or in part
The polypeptide of protein integrates the mammalian cell expression vector for encoding their polynucleotides as antigen.Exempted from using antigen
After epidemic disease animal, from through immunity animal adaptive immune cell and merge myeloma cell to obtain hybridoma.Then from hybridization
Knurl culture collects antibody.Finally can by using GFI1B albumen for being used as antigen or part thereof to obtain antibody reality
Apply antigentic specificity purifying to obtain the monoclonal antibody for GFI1B albumen.Polyclonal antibody can as follows be prepared：With with it is upper
Literary identical antigen-immunized animal, from the animal collect blood sample through immunity, isolates serum from blood, then uses
Above-mentioned antigen implements antigentic specificity purifying to serum.Can be by the antibody that obtained with ferment treatment or anti-by using what is obtained
The sequence information of body is obtaining antibody fragment.
The combination of label and antibody or its fragment can be implemented by method as commonly known in the art.For example, may be used
With following fluorescent marker protein or peptide：Clean protein or peptide with phosphate buffer, addition DMSO, buffer, etc. standard
Standby dyestuff, then mixed solution, places 10 minutes then at room temperature.In addition, mark can commodity in use labelling kit, it is all
Such as biotin labeling reagent box, such as biotin labeling reagent box-NH2, biotin labeling reagent box-SH
(DojindoLaboratories)；Alkali phosphatase enzyme mark kit such as alkali phosphatase enzyme mark kit-NH2, alkaline phosphorus
Sour enzyme labelling kit-SH (Dojindo Laboratories)；Peroxidase labelling kit such as peroxidase mark
Note kit-NH2, peroxidase labelling kit-NH2 (Dojindo Laboratories)；Phycobniliprotein labelled reagent
Box such as phycobniliprotein labelling kit-NH2, phycobniliprotein labelling kit-SH, B- phycoerythrin labelling kit-NH2,
B- phycoerythrin labelling kit-SH, R-PE labelling kit-NH2, R-PE labelling kit SH
(DojindoLaboratories)；Fluorescent labeling reagent box such as fluorescein labelling kit-NH2, HiLyte Fluor (TM)
555 labelling kit-NH2, the labelling kit-NH2 of HiLyte Fluor (TM) 647 (Dojindo Laboratories)；And
DyLight 547 and DyLight647 (Techno Chemical Corp.), Zenon (TM), Alexa Fluor (TM) antibody
Labelling kit, Qdot (TM) antibody labeling kit (Invitrogen Corporation) and EZ- label protein marks
Note kit (Funakoshi Corporation).For correct labeling, it is possible to use suitable instrument come detect through mark
Antibody or its fragment.
As the sample of the detection product according to the present invention, it is possible to use the tissue sample for for example obtaining from biopsy experimenter
Or fluid.Sample is not particularly limited, as long as it is suitable to the measure of the present invention；For example, it can include tissue, blood, blood plasma,
Serum, lymph liquid, urine, serous cavity liquid, spinal fluid, synovia, aqueous humor, tear, saliva or its fraction or treated material
Material.
In specific embodiments of the present invention, the sample source is in blood.
Further, the product of the quantitative GFI1B genes or GFI1B albumen can be detection GFI1B genes or GFI1B eggs
White reagent, can also be kit, chip, test paper comprising the reagent etc., or using the high pass of the reagent
Amount microarray dataset.
Present invention also offers a kind of instrument of diagnosis Bgudd-Chiari Syndrome, the instrument can detect GFI1B.
Further, the instrument being capable of GFI1B gene expression amounts in detection sample.
Further, the instrument include can quantitatively in sample GFI1B gene mRNAs reagent, and/or being capable of quantitative sample
The reagent of GFI1B albumen in product.
Further, it is described can quantitatively the reagent of GFI1B gene mRNAs be the spy used in real-time quantitative PCR in sample
The primer of different amplification GFI1B genes, the primer sequence is as shown in SEQ ID NO.1 and SEQ ID NO.2.It is described can be quantitative
The reagent of GFI1B albumen includes the antibody combined with GFI1B protein-specifics in sample.
Further, the instrument of the diagnosis Bgudd-Chiari Syndrome includes but is not limited to chip, kit, test paper or high flux
Microarray dataset；High-flux sequence platform is a kind of instrument of special diagnosis Bgudd-Chiari Syndrome, with high throughput sequencing technologies
Development, will become to the structure of the gene expression profile of a people and very easily work.By contrasting Disease and normal person
The gene expression profile of group, the exception for easily analyzing which gene is related to disease.Therefore, know in high-flux sequence
The exception of the GFI1B genes purposes for falling within GFI1B related to Bgudd-Chiari Syndrome, equally within protection scope of the present invention.
The amino acid that anti-GFI1B antibody or its fragment used in detection product, the diagnostic tool of the present invention is recognized
Number is not particularly limited, as long as antibody can be with reference to GFI1B.
Present invention also offers a kind of method of diagnosis Bgudd-Chiari Syndrome, methods described comprises the steps：
(1) sample of experimenter is obtained；
(2) expression of GFI1B genes or albumen in Samples subjects is detected；
(3) whether the expression of the GFI1B genes for measuring or albumen is associated with the illness of experimenter.
(4) compared with normal control, the expression of GFI1B genes or albumen is raised, then the experimenter is judged and has
Tendency with Bgudd-Chiari Syndrome or suffered from Bgudd-Chiari Syndrome, or Bgudd-Chiari Syndrome patient be judged as recurrence or
Person Bgudd-Chiari Syndrome patient is judged as prognosis mala.
In the context of the present invention, " diagnosis Bgudd-Chiari Syndrome " includes judging whether experimenter adds synthesis with cloth
Levy, judge that experimenter whether there is the risk with Bgudd-Chiari Syndrome, judge whether Bgudd-Chiari Syndrome patient has been recurred, judged
The prognosis of Bgudd-Chiari Syndrome patient.
The advantages of the present invention：
The present invention's is found that a kind of molecular marker of diagnosis Bgudd-Chiari Syndrome, can be in cloth using the molecular marker
Plus the early stage that syndrome occurs can judge, there is provided the survival rate of patient.
Specific embodiment
With reference to embodiment, the present invention is further detailed explanation.Following examples be merely to illustrate the present invention and
It is not used in restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in embodiment, generally according to normal condition, for example
Sambrook et al., molecular cloning：Laboratory manual (New York:Cold Spring HarborLaboratory Press,
1989) condition described in, or according to the condition proposed by manufacturer.
Embodiment 1 screens difference expression gene
1st, research object and sample collection
(1) research object：3 Bgudd-Chiari Syndrome patients and 5 Healthy Volunteers.
(2) sample collection：Research object at least 12h on an empty stomach is required, in m seq 7：00～8：Under 00 room temperature, extract
10ml venous blood adds 3 times of volume erythrocyte cracked liquids in ethylenediamine tetra-acetic acid (EDTA) anticoagulant tube, and room temperature places 10 after mixing
Minute, 10,000rpm centrifugations 1 minute.Thoroughly inhale and abandon supernatant, collect leukocyte cell pellet.It is white thin per 100-200 μ l blood collections
Born of the same parents' precipitation adds 1ml TRIzol, and -80 DEG C frozen standby.
2nd, the acquisition of total serum IgE
Conventionally extract the total serum IgE in blood leucocyte using TRIzol.
3rd, RNA concentration and purity testing
NanoDrop1000 spectrophotometers detect RNA sample, the sample requirement of RNA-seq sequencings：OD260/OD280 is
1.8-2.2。
4th, the quality analysis (Bioanalyzer of Agilent Technologies 2100) of RNA sample
The Bioanalyzer of Agilent Technologies 2100 detect RNA sample quality, observation 28S rRNA and
18S rRNA master tapes substantially, without degraded, the sequencing cDNA library that meets that RNA Perfection Index is qualified, concentration reaches requirement build
Requirement, can be used for library construction and sequencing.
5th, high flux transcript profile sequencing
(1) RNA-seq reads positioning
First by low-quality read remove obtain clean read, then using TopHat v1.3.1 will clean fragment and
UCSC H.sapiens reference gene groups (hg19) is matched, the index of the advance structure of H.sapiens UCSC hg19 versions
Download from TopHat homepages, and as reference gene group, when being matched with genome using TopHat, it is allowed to each read (acquiescence
To 20) having multiple matching sites, most 2 mispairing.TopHat sets up possible according to exon region and GT-AG shear signals
Shearing site storehouse, navigates to the read for not navigating to genome on genome according to these shearing site storehouses.We use
The system default parameter of TopHat methods.
(2) transcript abundance assessment
By Cufflinks v1.0.3 process, Cufflinks v1.0.3 are by RNA-seq pieces for the read file for matching
Hop count mesh is standardized the relative abundance for calculating transcript.FPKM values refer to specific per matching in 1,000,000 sequencing fragments
The segment number of the exon region of gene 1kb length.The confidential interval of FPKM estimates is calculated by Bayesian inference method.
The GTF comment files of the reference that Cufflinks is used download (Homo_ from Ensembl databases
sapiens.GRCh37.63.gtf)。
(3) detection of difference expression gene
The original document matched by the Ensembl GTF files of download and by TopHat is transferred to Cuffdiff,
Cuffdiff re-evaluates the gene expression abundance of the transcript listed in GTF files using original matching files, detects difference table
Reach.There was only q values ＜ 0.01 in Cuffidff outputs, test shows to be considered as successfully more just differential expression.
6th, result
Gene expression difference relatively in Bgudd-Chiari Syndrome patient and normal human blood, has 405 difference expression genes,
317 rises, 88 downwards.Wherein, compared with normal person, GFI1B genes up-regulated in Bgudd-Chiari Syndrome blood samples of patients,
MRNA relative expression quantities are 6.23 ± 1.51, and difference has statistical significance (P<0.05).
The difference expression gene that the checking of the large sample of embodiment 2 is filtered out
GFI1B genes are selected to be verified.
1st, sample collection
The peripheral blood sample that method according to embodiment 1 collects Bgudd-Chiari Syndrome patient and normal population is each 50.
2nd, verified in mRNA level in-site
2.1 extract blood total serum IgE
Step is with embodiment 1.
2.2 reverse transcriptions
Reverse transcription uses Primescript 1stStrand cDNA synthesis kit kits, operating procedure is as follows
Carry out：
(1) following reaction liquid is added in microcentrifugal tube, as shown in table 1：
The reaction liquid of table 1
(2) 70 DEG C of incubation 5min, are rapidly cooled to 4 DEG C；
Following reaction reagent is added in microcentrifugal tube, reaction system is made：
The preparation of the reaction system of table 2
| Reagent | Dosage |
| 5x1st Strand Synthesis Buffer | 4.0μl |
| PrimeScript RTase | 1.0μl |
| RNase Inhibitor | 1.0μl |
| Rnase free dH2O | 4.0μl |
Gently shake, after quick centrifugation, 42 DEG C of reaction 1h, 70 DEG C of 10min terminating reactions, 4 DEG C of coolings, -20 DEG C of preservations.
Using SYBP Premix Ex TapTMII kits, are carried out in Eppendorf Real-time PCR analyzers,
Concrete operations are as follows：
(1) following PCR reactant liquors are prepared on ice：
The preparation of the PCR reactant liquors of table 3
Primer sequence design is as follows：
GFI1B genes：
5’-CTCAAGTGCCTTCCTCTA-3’(SEQ ID NO.1)；
5’-CAGCTCTGGTCTACTCTT-3’(SEQ ID NO.2)
β-actin：
5’-GTGGGGCGCCCCAGGCACCA-3’(SEQ ID NO.3)；
5’-CTCCTTAATGTCACGCACGATTT-3’(SEQ ID NO.4)
(2) machine on, performs following programs：95 DEG C of denaturations 5min；95 DEG C of denaturation 15s.60 DEG C of annealing 15s, 72 DEG C of extensions
20s, totally 35 circulations.
As a result Bian relative quantification methods, formula 2-△△ctCalculate.Experiment is repeated 3 times.
△ ct=ct (A)-ct (β-actin)
△ △ ct=△ ct (experimental group)-△ ct (control group)
As a result show, compared with normal person, GFI1B genes up-regulated in Bgudd-Chiari Syndrome blood samples of patients, mRNA phases
It is 5.87 ± 1.25 to expression, difference has statistical significance (P<0.05).
The preparation of the Bgudd-Chiari Syndrome diagnostic kit of embodiment 2
According to GFI1B genes and the correlation of Bgudd-Chiari Syndrome, can be examined by detecting the expression of GFI1B genes
Disconnected Bgudd-Chiari Syndrome, accordingly the invention provides a kind of reagent that Bgudd-Chiari Syndrome is diagnosed based on detection GFI1B gene expressions
Box, the component in the diagnostic kit is as follows：SYBR Green PCR systems；Amplification GFI1B genes and β-
The primer pair of actin genes.The forward primer sequence of amplification GFI1B genes is 5 '-CTCAAGTGCCTTCCTCTA-3 ', reversely
Primer sequence is 5 '-CAGCTCTGGTCTACTCTT-3 '；Amplification β-actin forward primer sequence be 5 '-
GTGGGGCGCCCCAGGCACCA-3 ', reverse primer sequences are 5 '-CTCCTTAATGTCACGCACGATTT-3 '.SYBR
Green PCRs system includes PCR buffer solutions, dNTPs, SYBR Green fluorescent dyes.PCR buffer components
For：25mM KCL, 2.5mM MgCL2、200mM(NH4)2SO4。
The explanation of above-described embodiment is only intended to understand the method for the present invention and its core concept.It should be pointed out that for this
For the those of ordinary skill in field, under the premise without departing from the principles of the invention, some improvement can also be carried out to the present invention
And modification, these are improved and modification also will be fallen into the protection domain of the claims in the present invention.
SEQUENCE LISTING
<110>Beijing Yang Shen biology information technologies Co., Ltd
<120>A kind of Bgudd-Chiari Syndrome diagnostic tool
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 18
<212> DNA
<213>Artificial sequence
<400> 1
ctcaagtgcc ttcctcta 18
<210> 2
<211> 18
<212> DNA
<213>Artificial sequence
<400> 2
cagctctggt ctactctt 18
<210> 3
<211> 20
<212> DNA
<213>Artificial sequence
<400> 3
gtggggcgcc ccaggcacca 20
<210> 4
<211> 23
<212> DNA
<213>Artificial sequence
<400> 4
ctccttaatg tcacgcacga ttt 23
Claims (10)
1. application of the product of GFI1B in diagnosis Bgudd-Chiari Syndrome instrument is prepared is detected.
2. application according to claim 1, it is characterised in that the product of the detection GFI1B is included in detection sample
The product of GFI1B gene expression amounts.
3. application according to claim 2, it is characterised in that the product of the detection GFI1B includes being capable of quantitative sample
The product of middle GFI1B gene mRNAs, and/or can quantitatively in sample GFI1B albumen product.
4. application according to claim 3, it is characterised in that it is described can quantitatively in sample GFI1B gene mRNAs examination
Agent include real-time quantitative PCR used in specific amplified GFI1B genes primer, the primer sequence such as SEQ ID NO.1 with
Shown in SEQ ID NO.2；It is described can quantitatively the reagent of GFI1B albumen includes the anti-of specific binding GFI1B albumen in sample
Body.
5. the application according to any one of claim 2-4, it is characterised in that the sample source is in blood.
6. a kind of instrument of diagnosis Bgudd-Chiari Syndrome, it is characterised in that the instrument include can in detection sample GFI1B work
Tool.
7. instrument according to claim 6, it is characterised in that the instrument includes being capable of GFI1B genes in detection sample
The instrument of expression.
8. instrument according to claim 7, it is characterised in that the instrument includes being capable of quantitative GFI1B in sample
The reagent of gene mRNA, and/or can quantitatively in sample GFI1B albumen reagent.
9. instrument according to claim 8, it is characterised in that it is described can quantitatively in sample GFI1B gene mRNAs examination
Agent is the primer of the specific amplified GFI1B genes used in real-time quantitative PCR, the primer sequence such as SEQ ID NO.1 and SEQ
Shown in ID NO.2；It is described can quantitatively the reagent of GFI1B albumen includes the antibody of specific binding GFI1B albumen in sample.
10. the application according to any one of claim 6-9, it is characterised in that the sample source is in blood.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611222906.3A CN106636408A (en) | 2016-12-27 | 2016-12-27 | Budd-Chiari syndrome diagnosis tool |
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| CN201611222906.3A CN106636408A (en) | 2016-12-27 | 2016-12-27 | Budd-Chiari syndrome diagnosis tool |
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ID=58832685
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Citations (3)
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| CN106110297A (en) * | 2016-07-05 | 2016-11-16 | 南通大学 | The application of GFI1 truncate |
| WO2016182904A1 (en) * | 2015-05-08 | 2016-11-17 | President And Fellows Of Harvard College | Targeted selection of patients for treatment with cortistatin derivatives |
-
2016
- 2016-12-27 CN CN201611222906.3A patent/CN106636408A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101631861A (en) * | 2006-12-08 | 2010-01-20 | 奥斯瑞根公司 | As the miR-16 regulatory gene and the approach for the treatment of the target of intervening |
| WO2016182904A1 (en) * | 2015-05-08 | 2016-11-17 | President And Fellows Of Harvard College | Targeted selection of patients for treatment with cortistatin derivatives |
| CN106110297A (en) * | 2016-07-05 | 2016-11-16 | 南通大学 | The application of GFI1 truncate |
Non-Patent Citations (1)
| Title |
|---|
| 刘蕊，等: "《喉鳞癌组织与相邻正常黏膜的基因表达谱差异》", 《山东大学耳鼻喉眼学报》 * |
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