CN106755427A - Applications of the OTOF in Bgudd-Chiari Syndrome diagnosis - Google Patents
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- CN106755427A CN106755427A CN201611222917.1A CN201611222917A CN106755427A CN 106755427 A CN106755427 A CN 106755427A CN 201611222917 A CN201611222917 A CN 201611222917A CN 106755427 A CN106755427 A CN 106755427A
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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
-
- 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
Abstract
The invention discloses purposes of the OTOF in Bgudd-Chiari Syndrome diagnostic tool is prepared.The present invention is tested by the PCR of high-flux sequence and large sample, prove that expression of the OTOF genes in the blood of Bgudd-Chiari Syndrome patient is lowered, based on this, the present invention develops a kind of Bgudd-Chiari Syndrome diagnostic tool, the diagnostic tool by detecting OTOF gene expression amounts by obtain diagnostic result.Achievement in research of the invention provides a kind of new diagnosis thinking clinically to diagnose Bgudd-Chiari Syndrome.
Description
Technical field
The present invention relates to medical diagnosis on disease field, more particularly it relates to OTOF answering in Bgudd-Chiari Syndrome diagnosis
With.
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 lesion 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 types 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 diagnosis marker of Bgudd-Chiari Syndrome, the present invention is experimentally confirmed and suffers from Bgudd-Chiari Syndrome
OTOF gene mRNAs content is substantially less than normal population in the blood of person, thus the differential expression of OTOF genes property make its into
To can be used to diagnose the molecular marker whether subject suffers from Bgudd-Chiari Syndrome.
Specifically, the invention provides the application of the product in Bgudd-Chiari Syndrome diagnostic tool is prepared of detection OTOF.
Further, the product of the detection OTOF includes the product of OTOF gene expression amounts in detection sample.
Further, the product of the detection OTOF includes quantifying the product of OTOF gene mRNAs in sample, and/or
The product of OTOF albumen in sample can be quantified.
The product of OTOF gene mRNAs can be based on being sent out using the known method of nucleic acid molecules in quantitative sample of the 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 being prepared from biomaterial
Expect the gene of nucleic acid, then expand it to obtain using the primer for being designed for amplification 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 detected.
The product that OTOF gene mRNAs in sample can be quantified includes the specific amplified used in real-time quantitative PCR
The primer of OTOF 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 and prepared containing the gene for expecting nucleotide sequence from biomaterial, and expect that the primer of nucleotide sequence expands using amplification is designed for
Increase it to prepare.
The product of OTOF albumen can be based on playing its function using the known method of antibody in quantitative sample of the invention：
For example, can be including ELISA, radioimmunoassay, immunohistochemical method, western blot etc..
The product of OTOF albumen includes the antibody or its fragment of specific binding OTOF albumen in quantitative sample of the invention.
Can be using the antibody of any structure, size, immunoglobulin class, origin etc. or its fragment, as long as it combines target protein
.The antibody or its fragment that detection product of the invention includes can be monoclonals or polyclonal.Antibody fragment refers to
Retain peptide of the antibody to an antibody part (Partial Fragment) of the binding activity of antigen or containing an antibody part.Antibody fragment can
With including F (ab ')2, Fab ', Fab, scFv (scFv), disulphide bonding Fv (dsFv) or its polymer, dimerization V
Area's (double antibody) or the peptide containing CDR.The product of OTOF albumen can include encoding antibody or volume in quantitative sample of the invention
The nucleic acid of the separation of the amino acid sequence of code antibody fragment, the carrier comprising the nucleic acid, and carry the cell of the carrier.
Antibody can be obtained by well known to a person skilled in the art method.For example, prepare retaining 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 by immune animal adaptive immune cell and merging myeloma cell to obtain hybridoma.Then from hybridization
Knurl culture collects antibody.The antibody for obtaining can finally be implemented by using OTOF albumen for being used as antigen or part thereof
Antigentic specificity purifies to obtain the monoclonal antibody for OTOF albumen.Polyclonal antibody can as follows be prepared：With with above
Identical antigen-immunized animal, blood sample is collected from by immune animal, serum is isolated from blood, then using upper
State antigen and implement antigentic specificity purifying to serum.Can be by the antibody obtained with ferment treatment or the antibody by using acquisition
Sequence information obtain antibody fragment.
The combination of label and antibody or its fragment can be implemented by method as commonly known in the art.For example, can
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 be used the labelling kit of commercialization, 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 kits-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 by mark
Antibody or its fragment.
As the sample according to detection product of the invention, it is possible to use the tissue sample for for example being obtained from biopsy subject
Or fluid.Sample is not particularly limited, as long as it is suitable to measure of the invention；For example, it can include tissue, blood, blood plasma,
Serum, lymph, 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 OTOF genes or OTOF albumen can be detection OTOF genes or OTOF albumen
Reagent, can also be the kit comprising the reagent, chip, test paper etc., or using the reagent high pass measure
Sequence platform.
Present invention also offers a kind of instrument for diagnosing Bgudd-Chiari Syndrome, the instrument can detect OTOF.
Further, the instrument can detect OTOF gene expression amounts in sample.
Further, the instrument includes quantifying the reagent of OTOF gene mRNAs in sample, and/or can quantify sample
The reagent of middle OTOF albumen.
Further, the reagent that can quantify OTOF gene mRNAs in sample is the spy used in real-time quantitative PCR
The primer of different amplification OTOF genes, the primer sequence is as shown in SEQ ID NO.1 and SEQ ID NO.2.It is described to quantify
The reagent of OTOF albumen includes the antibody combined with OTOF 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 be as very easily working to a structure for the gene expression profile of people.By contrasting Disease and normal person
The gene expression profile of group, the exception for easily analyzing which gene is related to disease.Therefore, OTOF is known in high-flux sequence
The exception of the gene purposes for falling within OTOF related to Bgudd-Chiari Syndrome, equally within protection scope of the present invention.
The number of the amino acid that the anti-OTOF antibody or its fragment used in detection product of the invention, diagnostic tool are recognized
Mesh is not particularly limited, as long as antibody can combine OTOF.
Present invention also offers a kind of method for diagnosing Bgudd-Chiari Syndrome, methods described comprises the following steps：
(1) sample of subject is obtained；
(2) expression of OTOF genes or albumen in Samples subjects is detected；
(3) the OTOF genes or the expression of albumen that will be measured are associated with the whether ill of subject.
(4) compared with normal control, the expression reduction of OTOF genes or albumen, then the subject is judged has trouble
Have the tendency of Bgudd-Chiari Syndrome or suffered from Bgudd-Chiari Syndrome, or Bgudd-Chiari Syndrome patient be judged as recurrence or
Bgudd-Chiari Syndrome patient is judged as prognosis mala.
In the context of the present invention, " diagnosis Bgudd-Chiari Syndrome " includes judging whether subject adds synthesis with cloth
Levy, judge that subject 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：
It is of the invention to be found that a kind of molecular marker for diagnosing 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.
Brief description of the drawings
Fig. 1 displays detect the statistical chart of OTOF gene differential expression situations using QPCR in mRNA level in-site.
Specific embodiment
The present invention is further detailed explanation with reference to the accompanying drawings and examples.Following examples are merely to illustrate this
Invention rather than limitation the scope of the present invention.The experimental technique of unreceipted actual conditions in embodiment, generally according to conventional strip
Part, such as Sambrook et al., molecular cloning：Laboratory manual (New York:Cold Spring HarborLaboratory
Press, 1989) described in condition, 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：It is required that research object empty stomach at least 12h, in m seq 7：00~8：00 at room temperature, extracts
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, -80 DEG C freeze it is standby.
2nd, the acquisition of total serum IgE
Conventionally the total serum IgE in blood leucocyte is extracted 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 removal obtain clean read, then using TopHat v1.3.1 will clean fragment and
UCSC H.sapiens reference genes group (hg19) is matched, the index of the advance structure of H.sapiens UCSC hg19 editions
Downloaded from TopHat homepages, and as reference gene group, when being matched with genome using TopHat, it is allowed to each read (acquiescence
Sites, most 2 mispairing are matched to 20) there is multiple.TopHat sets up possible according to exon region and GT-AG shear signals
Shearing site storehouse, navigates on genome the read for not navigating to 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 treatment, 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 it is every 1,000,000 sequencing fragment in match it is specific
The segment number of exon region gene 1kb long.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
Cuffdiff is transferred to by the Ensembl GTF files of download and by the original document that TopHat is matched,
Cuffdiff re-evaluates the gene expression abundance of the transcript listed in GTF files using original matching files, detects difference table
Reach.The only q values ＜ 0.01 in Cuffidff outputs, test display is considered as successfully more just differential expression.
6th, result
Compare the gene expression difference in Bgudd-Chiari Syndrome patient and normal human blood, have 405 difference expression genes,
317 rises, 88 downwards.Wherein, compared with normal person, OTOF genes express downward in Bgudd-Chiari Syndrome blood samples of patients,
MRNA relative expression quantities are 0.31 ± 0.14, 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
Selection OTOF genes are verified.
1st, sample collection
Method according to embodiment 1 collects each 50 of the peripheral blood sample of Bgudd-Chiari Syndrome patient and normal population.
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
| Reagent | Dosage |
| RNA | 2.0μg |
| dNTP | 1.0μl |
| Oligo(dT) | 2.0μl |
| Rnase free dH2O | Add to 10.0 μ l |
(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 reaction solutions are being prepared on ice：
The preparation of the PCR reaction solutions of table 3
| Reagent | Dosage |
| SYBR | 10.0μl |
| Forward primer | 1.0μl |
| Reverse primer | 1.0μl |
| cDNA | 2.0μl |
| ddH2O | 6.0μl |
| Total amount | 20.0μl |
Primer sequence design is as follows：
OTOF genes：
5’-GAAGGTGGTAGAGGAGAG-3’(SEQ ID NO.1)；
5’-AGGCTGGTCTTGATGATA-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 predegeneration 5min；95 DEG C of denaturation 15s.60 DEG C of annealing 20s, 72 DEG C of extensions
20s, totally 42 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)
Result as shown in figure 1, compared with normal control population, the mRNA water of OTOF genes in Bgudd-Chiari Syndrome blood samples of patients
Dawn aobvious downward, difference has statistical significance (P<0.05).
The preparation of the Bgudd-Chiari Syndrome diagnostic kit of embodiment 2
According to OTOF genes and the correlation of Bgudd-Chiari Syndrome, can be diagnosed by detecting the expression of OTOF genes
Bgudd-Chiari Syndrome, diagnoses the kit of Bgudd-Chiari Syndrome the invention provides a kind of based on detection OTOF gene expressions accordingly,
Component in the diagnostic kit is as follows：SYBR Green PCR systems；Amplification OTOF genes and β-actin bases
The primer pair of cause.The forward primer sequence for expanding OTOF genes is 5 '-GAAGGTGGTAGAGGAGAG-3 ', reverse primer sequences
It is 5 '-AGGCTGGTCTTGATGATA-3 '；The forward primer sequence of amplification β-actin for 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 will be also fallen into the protection domain of the claims in the present invention.
SEQUENCE LISTING
<110>Beijing Yang Shen biology information technologies Co., Ltd
<120>Applications of the OTOF in Bgudd-Chiari Syndrome diagnosis
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 18
<212> DNA
<213>Artificial sequence
<400> 1
gaaggtggta gaggagag 18
<210> 2
<211> 18
<212> DNA
<213>Artificial sequence
<400> 2
aggctggtct tgatgata 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 detection OTOF in diagnosis Bgudd-Chiari Syndrome instrument is prepared.
2. application according to claim 1, it is characterised in that the product of the detection OTOF includes OTOF in detection sample
The product of gene expression amount.
3. application according to claim 2, it is characterised in that during the product of the detection OTOF includes to quantify sample
The product of OTOF gene mRNAs, and/or the product of OTOF albumen in sample can be quantified.
4. application according to claim 3, it is characterised in that the reagent that OTOF gene mRNAs in sample can be quantified
The primer of the specific amplified OTOF genes including being used in real-time quantitative PCR, the primer sequence such as SEQ ID NO.1 and SEQ
Shown in ID NO.2；The reagent that OTOF albumen in sample can be quantified includes the antibody of specific binding OTOF albumen.
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 for diagnosing Bgudd-Chiari Syndrome, it is characterised in that the instrument includes that the work of OTOF in sample can be detected
Tool.
7. instrument according to claim 6, it is characterised in that the instrument includes that OTOF gene tables in sample can be detected
Up to the instrument of amount.
8. instrument according to claim 7, it is characterised in that the instrument includes that OTOF bases in sample can be quantified
Because of the reagent of mRNA, and/or the reagent that OTOF albumen in sample can be quantified.
9. instrument according to claim 8, it is characterised in that the reagent that OTOF gene mRNAs in sample can be quantified
It is the primer of the specific amplified OTOF genes used in real-time quantitative PCR, the primer sequence such as SEQ ID NO.1 and SEQ ID
Shown in NO.2；The reagent that OTOF albumen in sample can be quantified includes the antibody of specific binding OTOF albumen.
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 |
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| CN201611222917.1A CN106755427A (en) | 2016-12-27 | 2016-12-27 | Applications of the OTOF in Bgudd-Chiari Syndrome diagnosis |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611222917.1A CN106755427A (en) | 2016-12-27 | 2016-12-27 | Applications of the OTOF in Bgudd-Chiari Syndrome diagnosis |
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| Publication Number | Publication Date |
|---|---|
| CN106755427A true CN106755427A (en) | 2017-05-31 |
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| CN201611222917.1A Pending CN106755427A (en) | 2016-12-27 | 2016-12-27 | Applications of the OTOF in Bgudd-Chiari Syndrome diagnosis |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2306191A1 (en) * | 2009-09-10 | 2011-04-06 | Miltenyi Biotec GmbH | Use of CD154 for the identification and separation of non-regulatory T-cells from a mixture of regulatory T-cells |
| WO2011159328A1 (en) * | 2010-06-16 | 2011-12-22 | Dynavax Technologies Corporation | Methods of treatment using tlr7 and/or tlr9 inhibitors |
-
2016
- 2016-12-27 CN CN201611222917.1A patent/CN106755427A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2306191A1 (en) * | 2009-09-10 | 2011-04-06 | Miltenyi Biotec GmbH | Use of CD154 for the identification and separation of non-regulatory T-cells from a mixture of regulatory T-cells |
| WO2011159328A1 (en) * | 2010-06-16 | 2011-12-22 | Dynavax Technologies Corporation | Methods of treatment using tlr7 and/or tlr9 inhibitors |
Non-Patent Citations (2)
| Title |
|---|
| 徐华英: "《彩色多普勒超声影像对布加综合征的诊断价值》", 《实用中西医结合临床》 * |
| 许伟，等: "《布-加综合征患者血清CA125、CA19-9的临床意义》", 《介入放射学杂志》 * |
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