CN106191291A - Rs1042658 application in predicting heating companion's thrombocytopenic syndromes mortality risk that new Bunyavirus causes - Google Patents
Rs1042658 application in predicting heating companion's thrombocytopenic syndromes mortality risk that new Bunyavirus causes Download PDFInfo
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- 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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- 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
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention discloses rs1042658 application in predicting heating companion's thrombocytopenic syndromes mortality risk that new Bunyavirus causes.The technical scheme that the present invention is protected is that in detection human genome, the polymorphism of rs1042658 or the material of genotype accompany the application in thrombocytopenic syndromes height mortality risk patient and the application in the product preparing heating companion's thrombocytopenic syndromes death risk that the new Bunyavirus of forecast China people causes in the heating that the preparation new Bunyavirus of examination Chinese causes.The material of the polymorphism of detection rs1042658 or genotype can be united, to other material (single nucleotide polymorphism or the material of genotype that thrombocytopenic syndromes death risk is relevant is accompanied in the heating caused as detected other Bunyavirus new with Chinese), heating companion's thrombocytopenic syndromes death risk that the new Bunyavirus of forecast China people causes.
Description
Technical field
The present invention relates to rs1042658 in biological technical field and predicting that platelet is accompanied in the heating that new Bunyavirus causes
Reduce the application in syndrome mortality risk.
Background technology
Heating companion's thrombocytopenic syndromes is a kind of emerging infectious disease being found first in rural areas in our country.Cause of disease
Body is a kind of new virus in bunyaviridae Phlebovirus, entitled new Bunyavirus.After being infected by this virus, warp
Cross the immunoreation of body self, still have the infected's morbidity of about 12% and clinical symptoms spectrum widely occurs, even may be used
Death, the inherited genetic factors of prompting individuality can be caused to play an important role in this disease.Identify that what new Bunyavirus caused sends out
The tumor susceptibility gene of heat companion's thrombocytopenic syndromes death contributes to predicting that heating companion's platelet that new Bunyavirus causes subtracts
The individual risk of few syndrome patient's death and group risk, and contribute to illustrating the pathogenesis relevant to this disease.Grain is thin
Born of the same parents' colony stimulating factor (G-CSF), is a kind of important hemopoietic growth factor, ties up human peripheral blood granulocyte quantity and function
Hold and play very pivotal role.Be related at present G-CSF and Peripheral blood cells relation and with the phase of some diseases
The existing more research report of closing property.Single nucleotide polymorphism (single nucleotide polymorphism, SNP) is to deposit
It is in the genomic DNA of a certain (a bit) colony, normal individual the sequence difference of single base, is that the restricted enzyme action length that continues is polymorphic
Third generation genetic marker after property and microsatellite.
Summary of the invention
The technical problem to be solved is how to predict heating companion's thrombocytopenia that new Bunyavirus causes
Syndrome mortality risk.
For solving above-mentioned technical problem, present invention firstly provides following arbitrary purposes:
A1) in detection human genome, the polymorphism of rs1042658 or the material of genotype are sick at preparation prediction Xin Buniya
Application in heating companion's thrombocytopenic syndromes mortality risk product that poison causes；
A2) in detection human genome, the polymorphism of rs1042658 or the material of genotype are sick at preparation examination Xin Buniya
Application in the heating companion thrombocytopenic syndromes height mortality risk patient product that poison causes；
A3) in detection human genome, the polymorphism of rs1042658 or the material of genotype detect and Xin Buniya in preparation
Application in the product of the single nucleotide polymorphism that heating companion's thrombocytopenic syndromes mortality risk that virus causes is correlated with；
A4) in detection human genome, the polymorphism of rs1042658 or the material of genotype are identified or auxiliary qualification in preparation
The product of the single nucleotide polymorphism that thrombocytopenic syndromes mortality risk is relevant is accompanied in the heating caused to new Bunyavirus
In application；
B1) in human genome, in preparation, the polymorphism of rs1042658 or genotype predict that what new Bunyavirus caused sends out
Application in heat companion's thrombocytopenic syndromes death risk product；
B2) in human genome the polymorphism of rs1042658 or genotype sending out that the preparation new Bunyavirus of examination causes
Application in heat companion thrombocytopenic syndromes height mortality risk patient product；
B3) in detection human genome, the polymorphism of rs1042658 or the material of genotype are predicting that new Bunyavirus draws
Application in the heating companion's thrombocytopenic syndromes mortality risk risen；
B4) in detection human genome, the polymorphism of rs1042658 or the material of genotype draw at the new Bunyavirus of examination
Application in the heating companion thrombocytopenic syndromes height mortality risk patient risen；
B5) in detection human genome, the polymorphism of rs1042658 or the material of genotype are detecting and new Bunyavirus
Application in the single nucleotide polymorphism that the heating companion's thrombocytopenic syndromes mortality risk caused is correlated with；
B6) in detection human genome, the polymorphism of rs1042658 or the material of genotype are being identified or are being assisted qualification with new
Application in the single nucleotide polymorphism that heating companion's thrombocytopenic syndromes mortality risk that Bunyavirus causes is correlated with；
B7) in human genome, polymorphism or the genotype of rs1042658 are predicting that the heating that new Bunyavirus causes is accompanied
Application in thrombocytopenic syndromes death risk；
B8) the heating companion that in human genome, the polymorphism of rs1042658 or genotype cause at the new Bunyavirus of examination
Application in thrombocytopenic syndromes height mortality risk patient.
Rs1042658 is the SNP site of two equipotential polymorphisms on human chromosome, this variation be conversion (C/T,
It is then G/A on its complementary strand).Described rs1042658 genotype is CC, CT or TT.Described CC be rs1042658 site be C
Homozygous, described TT be rs1042658 site be the homozygous of T, described CT be rs1042658 site be the heterozygous of C and T.
In described detection human genome, polymorphism or the genotype of rs1042658 specifically can be by detecting the nucleotide kind of rs1042658
Class determines.
In such use, in described detection human genome, the polymorphism of rs1042658 or the material of genotype can be amplification
The PCR primer of the genomic DNA fragment including rs1042658 and/or Single base extension primer.
In such use, heating companion's platelet that the individuality of described C/T and T/T genotype causes at new Bunyavirus subtracts
Heating companion's thrombocytopenic syndromes that ratio in syndrome patient's death colony causes at new Bunyavirus less than it less
Ratio in the non-dead colony of patient.
In such use, heating companion's thrombocytopenic syndromes that described new Bunyavirus causes can be that Chinese population is new
Heating companion's thrombocytopenic syndromes that Bunyavirus causes.
For solving above-mentioned technical problem, present invention also offers containing the polymorphism of rs1042658 in detection human genome
Or the product of the material of genotype.
The product of the material containing the polymorphism or genotype that detect rs1042658 in human genome provided by the present invention
Product, for a)-d) in any one product:
A) the monokaryon glycosides that thrombocytopenic syndromes mortality risk is relevant is accompanied in the heating that detection causes to new Bunyavirus
Acid polymorphism or the product of genotype；
B) heating companion's thrombocytopenic syndromes death wind that qualification and new Bunyavirus cause is identified or assists
Dangerous relevant single nucleotide polymorphism or the product of genotype；
C) the heating companion thrombocytopenic syndromes height mortality risk patient product that the new Bunyavirus of examination causes；
D) heating companion's thrombocytopenic syndromes death risk product that new Bunyavirus causes is predicted.
In the said goods, in described detection human genome, the polymorphism of rs1042658 or the material of genotype can be amplification
The PCR primer of the genomic DNA fragment including rs1042658 and/or Single base extension primer.
In the said goods, heating companion's thrombocytopenic syndromes that described new Bunyavirus causes can be that Chinese population is new
Heating companion's thrombocytopenic syndromes that Bunyavirus causes.
For solving above-mentioned technical problem, present invention also offers following M1) or method M2):
M1) method that thrombocytopenic syndromes mortality risk is accompanied in the heating that new Bunyavirus causes is predicted, including: inspection
Surveying the genotype in rs1042658 site in subject gene group to be measured, rs1042658 site is CC genotype object to be measured Xin Buni
Respectively higher than or candidate higher than rs1042658 site is the mortality risk of heating companion's thrombocytopenic syndromes that subvirus causes
CT genotype and the object to be measured of TT genotype；
The method of the heating companion thrombocytopenic syndromes height mortality risk patient that M2) the new Bunyavirus of examination causes,
Including: detecting the genotype in rs1042658 site in subject gene group to be measured, rs1042658 site is the to be measured of CC genotype
Object is or candidate is the heating companion thrombocytopenic syndromes height mortality risk patient that new Bunyavirus causes；
Rs1042658 site is that the object to be measured of CT genotype is or candidate is heating companion's thrombocytopenia that new Bunyavirus causes
Syndrome low mortality risk patient；Rs1042658 site is that the object to be measured of TT genotype is or candidate is new Bunyavirus
The heating companion thrombocytopenic syndromes low mortality risk patient caused.
In said method, detect the genotype in rs1042658 site in subject gene group to be measured and can use described detection
The polymorphism of rs1042658 or the material of genotype are carried out.
In said method, heating companion's thrombocytopenic syndromes that described new Bunyavirus causes can be that Chinese population is new
Heating companion's thrombocytopenic syndromes that Bunyavirus causes.
In the present invention, described PCR primer can be P1 and P2；Described P1 is following a1) to a4) in any one single stranded DNA:
A1) single stranded DNA shown in SEQ ID No.1 in sequence table；
A2) at a1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
A3) and a1) or a2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
A4) under strict conditions with a1) or a2) limit single stranded DNA hybridization single stranded DNA；
Described P2 can be following b1) to b4) in any one single stranded DNA:
B1) single stranded DNA shown in SEQ ID No.2 in sequence table；
B2) at b1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
B3) and b1) or b2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
B4) under strict conditions with b1) or b2) limit single stranded DNA hybridization single stranded DNA.
Described Single base extension primer can be following c1) to c4) in any one single stranded DNA:
C1) single stranded DNA shown in SEQ ID No.3 in sequence table；
C2) at c1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
C3) and c1) or c2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
C4) under strict conditions with c1) or c2) limit single stranded DNA hybridization single stranded DNA.
A2) described at a1) 5 ' ends and/or 3 ' ends add single stranded DNAs that one or several nucleotide obtain at SEQ
5 ' ends and/or the 3 ' ends of the single stranded DNA shown in ID No.1 add the single stranded DNA that one to ten nucleotide obtains.B2) described
B1) single stranded DNA that 5 ' ends and/or 3 ' hold one or several nucleotide of interpolation to obtain is at the strand shown in SEQ ID No.2
5 ' ends and/or the 3 ' ends of DNA add the single stranded DNA that one to ten nucleotide obtains.C2) described at c1) 5 ' ends and/or 3 ' ends
Adding the single stranded DNA that one or several nucleotide obtains is at 5 ' ends of the single stranded DNA shown in SEQ ID No.3 and/or 3 ' ends
Add the single stranded DNA that one to ten nucleotide obtains.
Term used herein " homogeneity " refers to the sequence similarity with native sequence nucleic acid." homogeneity " includes and this
Nucleotide sequence shown in bright SEQ ID No.1, SEQ ID No.2 or SEQ ID No.3 has 85% or higher, or 90%
Or higher, or the nucleotide sequence of 95% or higher homogeneity.Homogeneity can with the naked eye or computer software is evaluated.Make
With computer software, the homogeneity between two or more sequences can use percentage ratio (%) to represent, it can be used to evaluate phase
Close the homogeneity between sequence.
Described stringent condition is at 2 × SSC, in the solution of 0.1%SDS, hybridizes and washes film 2 times, every time at 68 DEG C
5min, again in 0.5 × SSC, in the solution of 0.1%SDS, hybridizes at 68 DEG C and washes film 2 times, each 15min；Or, 0.1 ×
SSPE (or 0.1 × SSC), 0.1%SDS solution in, hybridize under the conditions of 65 DEG C and wash film.
Above-mentioned more than 85% homogeneity, can be the homogeneity of 85%, 90% or more than 95%.
It is demonstrated experimentally that in the heating companion thrombocytopenic syndromes PATIENT POPULATION that new Bunyavirus causes,
The T equipotential of rs1042658 ratio in death group is less than its ratio in non-death group, has statistics poor
Different, the guarantor that T equipotential is heating companion's thrombocytopenic syndromes death that new Bunyavirus causes of rs1042658 is described
Protect equipotential；The C equipotential of rs1042658 ratio in death group is higher than its ratio in non-death group, has
Significant difference, illustrates that C equipotential is that the heating companion thrombocytopenic syndromes patient that the new Bunyavirus of risk equipotential causes is dead
The risk equipotential died.In the heating companion thrombocytopenic syndromes PATIENT POPULATION that new Bunyavirus causes, rs1042658 position
In three genotype of point, the individuality of C/T and T/T genotype ratio in death group is less than it in non-death group
In ratio, the individuality of C/C genotype ratio in death group be higher than its ratio in non-death group.Newly
In the heating companion thrombocytopenic syndromes PATIENT POPULATION that Bunyavirus causes, compared with the individuality carrying C/C genotype, take
Individual heating companion's thrombocytopenic syndromes mortality risk with C/T and T/T genotype reduces, odds ratio (Odds ratio,
OR) it is 0.46, P=0.018, illustrates that heating companion's platelet that rs1042658 pleomorphism site and new Bunyavirus cause subtracts
The generation of few syndrome mortality risk significantly associates.
In actual applications, the polymorphism of rs1042658 or the material of genotype will can be detected with other material (such as detection
The mononucleotide that thrombocytopenic syndromes mortality risk is relevant is accompanied in the heating that other Bunyavirus new to Chinese causes
Polymorphism or the material of genotype) be united preparation the new Bunyavirus of forecast China people cause heating companion platelet subtract
The product of few syndrome patient's mortality risk.
Wherein, in detection human genome, the polymorphism of rs1042658 or the material of genotype can be by following at least one
Kind of method determines the reagent needed for the polymorphism of rs1042658 or genotype and/or instrument: DNA sequencing, restriction fragment
Length polymorphism, single strand conformation polymorphism, denaturing high-performance chromatography, SNP chip, TaqMan probe technology and Sequenom
MassArray technology.Wherein, Sequenom MassArray technology is utilized to determine polymorphism or the genotype institute of rs1042658
The reagent needed and/or instrument include PCR primer to, extension primer based on single base extension, phosphatase be (such as shrimp alkalescence phosphorus
Acid enzyme (shrimp alkaline phosphatase, SAP)), resin, chip, MALDI-TOF (matrix-assisted
Laser desorption/ionization time of fligh, matrix solid-dispersion flight time mass spectrum)
And other reagent required for Sequenom MassArray technology and instrument；TaqMan probe technology is utilized to determine
Reagent needed for the polymorphism of rs1042658 or genotype and/or instrument include that TaqMan probe, PCR primer are to, quantitative PCR
Instrument and carry out gene type software (such as 7500 System SDS software) and TaqMan probe technology required for
Other reagent；SNP chip include chip based on nucleic acid hybridization reaction, chip based on single base extension, based on equipotential
The chip of gene-specific primer extension, the chip reacted based on " one-step method ", chip based on primer coupled reaction, base
In chip, the chip based on protein D NA association reaction of restriction enzyme reaction, and based on fluorescence molecule DNA association reaction
Chip.
Described product can be reagent or test kit, can be also the system being made up of with instrument reagent or test kit, as by drawing
Thing and the system of DNA sequencer composition, the system being made up of PCR reagent and DNA sequencing reagent and DNA sequencer, by TaqMan
Other examinations required for software that probe, PCR primer to, quantitative PCR apparatus and carry out gene type and TaqMan probe technology
Agent composition system, by PCR primer to, Single base extension primer, chip, PCR instrument and carry out gene type software and
Other reagent required for Sequenom MassArray technology and the system of instrument composition.
In one embodiment of the present of invention, Sequenom MassArray technology is used to determine the polymorphism of rs1042658
And genotype.Described PCR primer is to not having particular/special requirement in sequence, as long as the base including rs1042658 can be amplified
Because of group DNA fragmentation, the concretely single stranded DNA shown in SEQ ID No.1 and SEQ ID No.2 in sequence table.Described list
Base extends the single stranded DNA shown in SEQ ID No.3 in primer concretely sequence table.Sequenom MassArray technology institute
Other instruments needed are MassARRAY Nanodispenser RS1000 point sample instrument (SEQUENOM company).Sequenom
Chip required for MassArray technology is SpectroCHIP (Sequenom) chip.The described software tool carrying out gene type
Body is MALDI-TOF and TYPER 4.0 software (sequenom).
(heating companion's thrombocytopenia that new Bunyavirus causes is comprehensive at a sample from Chinese population for the present invention
Levy the non-dead colony of patient and new Bunyavirus causes heating companion thrombocytopenic syndromes death colony) in find
Rs1042658 is the monokaryon glycosides that the heating companion's thrombocytopenic syndromes mortality risk height caused to new Bunyavirus is relevant
Acid polymorphism.Can be by the material of the detection polymorphism of rs1042658 or genotype and other material (as that detect other with new cloth
The single nucleotide polymorphism or genotype that thrombocytopenic syndromes death risk is relevant is accompanied in the heating that Buddhist nun's subvirus causes
Material) be united preparation predict new Bunyavirus cause heating companion thrombocytopenic syndromes death risk
Product.
Detailed description of the invention
Being further described in detail the present invention below in conjunction with detailed description of the invention, the embodiment be given is only for explaining
The bright present invention rather than in order to limit the scope of the present invention.
Experimental technique in following embodiment, if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Embodiment 1 detects the foundation of the rs1042658 loci gene type method of G-CSF gene
1, the extraction of genomic DNA
Extract testing sample genomic DNA, carry out genotype detection with it for template.
2, the design of primer and synthesis
Use Sequenom company Genotyping Tools and MassARRAY Assay Design software design to be measured
The pcr amplification primer thing of SNP site and Single base extension primer, and transfer to biotech firm to synthesize.Detection G-CSF gene
The sequence of the specific primer pair of rs1042658 genotype is: forward primer (its entitled P1) 5 '-
ACGTTGGATGATTCCTCCTGTCTGCTCCCT-3 ' (the SEQ ID No.1 in sequence table)；Reverse primer (its entitled P2)
5 '-ACGTTGGATGGTGACTCTTTTTAGGGCCAG-3 ' (the SEQ ID No.2 in sequence table).Anti-for single base amplification
The extension primer sequence answered is 5 '-gggtACATTTGCCTTGCTGGA-3 ' (the SEQ ID No.3 in sequence table).
3, the foundation of the rs1042658 loci gene type method of detection G-CSF gene
3.1 PCR amplifications: PCR amplification is carried out in 384 orifice plates, and each reaction system cumulative volume is 5 μ L, prepares by table 1
PCR reaction system.
The component of table 1 each PCR reaction system
PCR response procedures is: 94 DEG C 4 minutes；94 DEG C 20 seconds, 56 DEG C 30 seconds, 72 DEG C 1 minute, 45 circulations；72 DEG C 3 points
Clock；4 DEG C of holdings.
3.2 PCR primer alkaline phosphatase treatment: after PCR reaction terminates, by PCR primer SAP (shrimp
Alkaline phosphatase, shrimp alkaline phosphotase) process, with remove system middle reaches from dNTPs, prepare SAP by table 2 anti-
Answer system.
Table 2 SAP reaction system
| Reagent | Volume (μ L) |
| Ultra-pure water | 1.53 |
| 10 × SAP buffer | 0.17 |
| SAP enzyme (1.7U/ μ l) | 0.3 |
| Cumulative volume | 2 |
Reaction condition is: 37 DEG C 40 minutes；85 DEG C 5 minutes；4 DEG C of maintenances.
3.3 Single base extensions: utilize 10 × single base extension buffer, single alkali after alkaline phosphatase treatment terminates
The extension primer of the single base amplification reaction shown in base extension enzyme (1.7U/ μ l) and SEQ ID No.3 carries out single base and prolongs
Stretch reaction.
Reaction condition is:
I.94 DEG C 30 seconds
II.94 DEG C 5 seconds
III.52 DEG C 5 seconds
IV.80 DEG C 5 seconds
V. III, 4 circulations are returned to
VI. II, 39 circulations are returned to
VII.72 DEG C 3 minutes
VII.4℃
3.4 resin purifications: Clean Resin resin is tiled in the resin plate of 6mg；Add 16 μ l water to extension products
In corresponding aperture；Being poured into by dried resin in extension products plate, sealer, slow speed vertical rotates 30 minutes, makes resin and reaction
Thing is fully contacted；Being centrifuged makes resin sink to bottom hole.
3.5 chip point samples: start MassARRAY Nanodispenser RS1000 point sample instrument (SEQUENOM company), will
Extension products after resin purification moves on 384 hole SpectroCHIP (Sequenom) chips (SEQUENOM company).
3.6 Mass Spectrometer Method: using MALDI-TOF to analyze the SpectroCHIP chip after point sample, testing result uses
TYPER 4.0 software (sequenom) typing also exports result.
Heating companion's blood that the rs1042658 site Bunyavirus new with Chinese of embodiment 2 G-CSF gene causes is little
Plate reduces the analysis of syndrome death susceptibility
The method set up by embodiment 1, to Chinese population, (heating companion's blood that the 430 new Bunyavirus of example cause is little
Plate reduces syndrome patient) the rs1042658 loci gene type of G-CSF gene analyzed.
1, object of study
Case group: the heating companion thrombocytopenic syndromes patient that the 430 new Bunyavirus of example cause, and all meet new cloth
The heating that Buddhist nun's subvirus causes is accompanied thrombocytopenic syndromes patient clinical diagnostic criteria and examines through new Bunyavirus nucleic acid
Survey and confirm.Wherein 48 people are dead.382 not dead example patients are classified as non-death group, 48 dead example patients are classified as
Death group.
All object of study are the Chinese han population of consanguinity-less relation.All object of study all endorsed informed consent
Book, collects population statistics and the medical history of individual by structurized questionnaire.This research obtains the human relations of hospital of PLA 154
The approval of reason committee.
2, the determination of the rs1042658 loci gene type of G-CSF gene
In crowd, the genotype in the rs1042658 site of G-CSF gene and the frequency distribution of equipotential are as shown in table 3.
Result shows, in all of object of study, in non-death group, the frequency of C equipotential is 55.5%, T equipotential
Frequency is 44.5%；In death group, the frequency of C equipotential is 67.7%, and the frequency of T equipotential is 32.3%.T equipotential is in death
The frequency ratio in case group frequency in non-death group is low, has significant difference, shows the T in rs1042658 site
Equipotential is the protection equipotential of heating companion's thrombocytopenic syndromes death that the new Bunyavirus of Chinese causes, and
The C equipotential of rs1042658 ratio in death group is higher than its ratio in non-death group, has statistics poor
Different, the risk equipotential that C equipotential is heating companion's thrombocytopenic syndromes death that the new Bunyavirus of Chinese causes is described.
In three genotype in all of object of study rs1042658 site, the individuality of C/C genotype is in death
Ratio in group is higher than its ratio in non-death group, and the individuality of C/T and T/T ratio in death group is respectively
Less than the individuality of corresponding gene type ratio in non-death group.In all of object of study, and carry C/C genotype
Individuality compare, carry heating companion's thrombocytopenic syndromes that the individual new Bunyavirus of C/T and T/T genotype causes dead
Occurrence risk of dying reduces, and OR value is 0.46 [95% confidence interval (95%CI)=(0.24-0.87)；P=0.018].
Result shows, it is possible to use heating companion's blood that the new Bunyavirus of rs1042658 site estimation Chinese causes is little
Plate reduces the mortality risk of syndrome patient.
Heating companion's platelet that the rs1042658 site Bunyavirus new with Chinese of table 3 G-CSF gene causes subtracts
The association analysis that few syndrome is dead
Note: OR, odds ratio (odds ratio)；CI, confidence interval (confidence interval).
aData are through logistic regression analysis, and carried out the correction of age, sex and underlying diseases.
bχ2Inspection.
Claims (10)
1. in detection human genome, in preparation, the polymorphism of rs1042658 or the material of genotype predict that new Bunyavirus causes
Heating companion thrombocytopenic syndromes mortality risk product in application.
2. in detection human genome, the polymorphism of rs1042658 or the material of genotype cause at the preparation new Bunyavirus of examination
Heating companion thrombocytopenic syndromes height mortality risk patient product in application.
3. in detection human genome, the polymorphism of rs1042658 or the material of genotype draw with new Bunyavirus in preparation detection
Application in the product of the single nucleotide polymorphism that the heating companion's thrombocytopenic syndromes mortality risk risen is correlated with.
4. in detection human genome, the polymorphism of rs1042658 or the material of genotype are identified in preparation or assist qualification and new cloth
Answering in the product of the single nucleotide polymorphism that heating companion's thrombocytopenic syndromes mortality risk that Buddhist nun's subvirus causes is correlated with
With.
The most following arbitrary application:
B1) in human genome, polymorphism or the genotype of rs1042658 predict, in preparation, the heating companion that new Bunyavirus causes
Application in thrombocytopenic syndromes death risk product；
B2) in human genome, polymorphism or the genotype of rs1042658 are preparing the heating companion that the new Bunyavirus of examination causes
Application in thrombocytopenic syndromes height mortality risk patient product；
B3) in detection human genome, the polymorphism of rs1042658 or the material of genotype are predicting what new Bunyavirus caused
Application in heating companion's thrombocytopenic syndromes mortality risk；
B4) detect what the polymorphism of rs1042658 or the material of genotype in human genome caused at the new Bunyavirus of examination
Application in heating companion thrombocytopenic syndromes height mortality risk patient；
B5) in detection human genome, the polymorphism of rs1042658 or the material of genotype cause with new Bunyavirus in detection
The relevant single nucleotide polymorphism of heating companion's thrombocytopenic syndromes mortality risk in application；
B6) in detection human genome, the polymorphism of rs1042658 or the material of genotype are being identified or are being assisted qualification and Xin Buni
Application in the single nucleotide polymorphism that heating companion's thrombocytopenic syndromes mortality risk that subvirus causes is correlated with；
B7) in human genome, polymorphism or the genotype of rs1042658 are predicting that heating companion's blood that new Bunyavirus causes is little
Plate reduces the application in syndrome patient's mortality risk；
B8) in human genome, polymorphism or the genotype of rs1042658 accompany blood little in the heating that the new Bunyavirus of examination causes
Plate reduces the application in syndrome height mortality risk patient.
6. contain the product of the material of the polymorphism of rs1042658 or genotype in detection human genome, for a)-d) in arbitrary
Kind product:
A) heating that detection causes to new Bunyavirus accompanies the mononucleotide that thrombocytopenic syndromes mortality risk is relevant many
State property or the product of genotype；
B) heating companion's thrombocytopenic syndromes death risk phase that qualification and new Bunyavirus cause is identified or assists
The single nucleotide polymorphism closed or the product of genotype；
C) the heating companion thrombocytopenic syndromes height mortality risk patient product that the new Bunyavirus of examination causes；
D) heating companion's thrombocytopenic syndromes death risk product that new Bunyavirus causes is predicted.
Product the most according to claim 6, it is characterised in that: the polymorphism of rs1042658 in described detection human genome
Or the material of genotype is PCR primer and/or the Single base extension expanding the genomic DNA fragment including rs1042658
Primer.
Product the most according to claim 7, it is characterised in that: described PCR primer is P1 and P2；Described P1 is following a1)
To a4) in any one single stranded DNA:
A1) single stranded DNA shown in SEQ ID No.1 in sequence table；
A2) at a1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
A3) and a1) or a2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
A4) under strict conditions with a1) or a2) limit single stranded DNA hybridization single stranded DNA；
Described P2 is following b1) to b4) in any one single stranded DNA:
B1) single stranded DNA shown in SEQ ID No.2 in sequence table；
B2) at b1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
B3) and b1) or b2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
B4) under strict conditions with b1) or b2) limit single stranded DNA hybridization single stranded DNA.
9. according to the product described in claim 7 or 8, it is characterised in that: described Single base extension primer is following c1) to c4)
In any one single stranded DNA:
C1) single stranded DNA shown in SEQ ID No.3 in sequence table；
C2) at c1) 5 ' ends and/or 3 ' ends add the single stranded DNAs that obtain of one or several nucleotide；
C3) and c1) or c2) single stranded DNA that limits has the single stranded DNA of the homogeneity of more than 85%；
C4) under strict conditions with c1) or c2) limit single stranded DNA hybridization single stranded DNA.
The most following M1) or method M2):
M1) method that thrombocytopenic syndromes mortality risk is accompanied in the heating that new Bunyavirus causes is predicted, including: detection is treated
Surveying the genotype in rs1042658 site in subject gene group, rs1042658 site is that CC genotype object to be measured Xin Buniya is sick
The mortality risk of heating companion's thrombocytopenic syndromes that poison causes is respectively higher than or candidate is CT base higher than rs1042658 site
Because of type and the object to be measured of TT genotype；
The method of the heating companion thrombocytopenic syndromes height mortality risk patient that M2) the new Bunyavirus of examination causes, including:
Detecting the genotype in rs1042658 site in subject gene group to be measured, rs1042658 site is that the object to be measured of CC genotype is
Or candidate is the heating companion thrombocytopenic syndromes height mortality risk patient that new Bunyavirus causes；Rs1042658 site
Object to be measured for CT genotype is or candidate is heating companion's low death of thrombocytopenic syndromes that new Bunyavirus causes
Risk patient；Rs1042658 site is that the object to be measured of TT genotype is or candidate is the heating companion that new Bunyavirus causes
Thrombocytopenic syndromes low mortality risk patient.
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