CN105861707B - IL-1 β gene pleiomorphism relevant to premature labor generation and its detection method - Google Patents
IL-1 β gene pleiomorphism relevant to premature labor generation and its detection method Download PDFInfo
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Abstract
The invention discloses IL-1 β gene pleiomorphism relevant to premature labor generation and its detection methods.The present invention provides the substances of the polymorphism in the site IL-1 β gene rs1143634 in detection newborn's genome or genotype to identify or assist to identify whether newborn is application in spontaneous pre-term.The present invention has studied correlation of the polymorphic site (G3953T (rs1143634)) of gene IL-1 β with premature labor neurological susceptibility, distinguish carry the site IL-1 β G3953T T allele, at least one T allelotype (CT+TT genotype) newborn occur SPTB (spontaneous pre-term) risk increase.
Description
Technical field
The present invention relates to field of biotechnology, more particularly to IL-1 β gene pleiomorphism relevant to premature labor generation and its inspection
Survey method.
Background technique
Premature labor (Preterm Birth, PTB) is to lead to neonatal morbidity and dead major reason, while and causing
Brain paralysis, hypoevolutism, retinopathy of prematurity, chronic lung disease and the sense of hearing and dysopia etc. occur for infant
An important factor for complication and sequelae.75% neonatal morbidity and death are related with premature labor.In the whole world, there are about 1500 every year
Ten thousand premature, and this number is constantly rising.It is shown according to World Health Organization's publication data, China premature in 2011
Disease incidence be 7.8%, have nearly 1,100,000 Premature Birth, birth is next in number only to India.Premature labor have become one it is worldwide
Hygienic issues.Existing epidemiological study shows that premature labor has apparent Familial aggregation phenomenon.And the disease incidence tool of premature labor
There is race and ethnic origin difference, the disease incidence of premature labor is Europe white twice in African American population.The studies above
Show that inherent cause plays an important role in spontaneous pre-term morbidity.Therefore, it is early to identify that the tumor susceptibility gene of premature labor facilitates prediction
The individual risk and group risk occurred is produced, and helps to illustrate pathogenesis relevant to this disease.
Gestation is a complicated physiology course, and inflammatory reaction plays an important role in premature labor and the morbidity of premature rupture of fetal membranes,
The main reason for being premature labor and premature rupture of fetal membranes generation.The gene of inflammatory cytokine plays important work in premature labor and premature rupture of fetal membranes
With being the research premature labor and good candidate gene of premature rupture of fetal membranes neurological susceptibility.It is well known that IL-1 is in immune and inflammatory reaction tune
Section aspect plays a significant role, and IL-1 α and IL-1 β are the cell factors with critical function, and wherein IL-1 β is considered scorching
The important regulatory factor of disease reaction, it is closely related with the proliferation of cell, differentiation, apoptosis.The gene cluster of coding IL-1 is located at 2q13-
On 21, length is about 430bp, the gene including coding IL-1 α, IL-1 β and IL-1RN.The SNPs of these genes is adjustable
The biological efficiency of IL-1 signal and then the occurrence risk for influencing disease.The SNPs of IL-1 gene cluster gene is with kinds cancer and slowly
The genetic predisposition of sexy infectious diseases is related, such as: the cancer of the esophagus, lung cancer, colon cancer, systemic loupus erythematosus, case-hardening
Disease, ulcerative colitis, chronic hepatitis C and type II diabetes periodontosis.The SNPs of IL-1 β gene can influence peripheral blood
The expression of IL-1 β in monocyte.The C+3953T polymorphic site (rs1143634) of IL-1 β gene is located at IL-1 β base
On 5th exon of cause, which has great importance to the adjusting of IL-1 β expression.In vitro experiment, L-
MinorAllele IL-1 β+3953*2 (the T equipotential base that the C of the rs1143634 polymorphic site of 1 β gene is formed after being replaced by T
Cause) it is related with the expression increase of IL-1 β.In European crowd, compared with C allele, rs1143634 polymorphic site
The level of T allele and higher blood glucose it is related, thus, the risk that the individual of carrying T allele suffers from diabetes is significant
Increase.
Summary of the invention
A purpose of the invention be to provide in detection Fetal genome the polymorphism in the site IL-1 β gene rs1143634 or
The purposes of allele or the substance of genotype.
The polymorphism or allele in the site IL-1 β gene rs1143634 in detection Fetal genome provided by the invention
Or the following purposes of the substance of genotype:
Detect the polymorphism in the site IL-1 β gene rs1143634 or the object of allele or genotype in Fetal genome
Matter is that spontaneous pre-term easily send out application in individual probability character predicting or assisting prediction fetus；
Or the polymorphism or allele or genotype in the site IL-1 β gene rs1143634 in detection Fetal genome
Substance is that spontaneous pre-term easily send out application in individual probability character product in preparation prediction or auxiliary prediction fetus；
Or the polymorphism or allele or genotype in the site IL-1 β gene rs1143634 in detection Fetal genome
Application of the substance in the risk character that spontaneous pre-term occurs for assessment or aided assessment fetus to be measured；
Or the polymorphism or allele or genotype in the site IL-1 β gene rs1143634 in detection Fetal genome
Application of the substance in the risk character product that spontaneous pre-term occurs for preparation assessment or aided assessment fetus to be measured.
Another object of the present invention provides the polymorphism in the site IL-1 β gene rs1143634 in detection newborn's genome
Or the different purposes of allele or the substance of genotype.
The polymorphism or equipotential base in the site IL-1 β gene rs1143634 in detection newborn's genome provided by the invention
The following purposes of the substance of cause or genotype:
Detect the polymorphism or allele or genotype in the site IL-1 β gene rs1143634 in newborn's genome
Substance is being identified or is assisting to identify newborn for the application in the probability character of spontaneous pre-term；
Or the polymorphism in the site IL-1 β gene rs1143634 or allele or genotype in detection newborn's genome
Substance identify or assist to identify newborn for the application in the probability character product of spontaneous pre-term in preparation.
In above-mentioned application, the genotype in the site rs1143634 is CC or CT；
The allele in the site rs1143634 is C or T；The site rs1143634 is located at human genome the 2nd
The 14th on 5th exon of the IL-1 β gene of chromosome；
And/or the nucleotide sequence of the IL-1 β gene is specially sequence 1.
3rd purpose of the invention is to provide following method.
The present invention provides following method 1) -3) in it is any:
Method 1): a method of prediction or auxiliary prediction fetus are that spontaneous pre-term easily send out individual probability, for following A or
B,
A includes the following steps: that the genotype in the site rs1143634 for the IL-1 β gene for detecting fetus to be measured is CC or CT,
The fetus to be measured of CC genotype is that spontaneous pre-term easily send out individual probability and is less than or the candidate fetus to be measured less than CT genotype；
B includes the following steps: that the allele in the site rs1143634 for the IL-1 β gene for detecting fetus to be measured is C or T,
The fetus to be measured that allele is C is that spontaneous pre-term easily sends out that individual probability is less than or the candidate allele that is less than is the to be measured of T
Fetus；
Method 2): a method of the risk character of spontaneous pre-term occurs for assessment or aided assessment fetus to be measured, be A or
B,
A includes the following steps: that the genotype in the site rs1143634 for the IL-1 β gene for detecting fetus to be measured is CC or CT,
The risk that spontaneous pre-term occurs for CC genotype fetus to be measured is less than or candidate is less than CT genotype fetus to be measured；
B includes the following steps: that the allele in the site rs1143634 for the IL-1 β gene for detecting fetus to be measured is C or T,
The risk that spontaneous pre-term occurs for the fetus to be measured of C allele is less than or the candidate fetus to be measured for being less than T allele；
Method 3): a method of it identifies or assists to identify the probability character that newborn is spontaneous pre-term, be following A
Or B,
A includes the following steps: to detect
CC or CT, CC genotype wait for that detecting new-born baby is that the probability of spontaneous pre-term is less than or candidate is less than CT genotype new life to be measured
Youngster；
B includes the following steps: to detect the allele to the site IL-1 β gene rs1143634 in detecting new-born baby genome
For C or T, C allele is less than to the probability that detecting new-born baby is spontaneous pre-term or candidate is less than the to be measured of T allele
Newborn.
In the above method,
Each sample genotype of the detection or the method for allele include the following steps:
A, the primer pair of the composition of single strand dna shown in the single stranded DNA shown in sequence 1 and sequence 2 is to each sample base
Because group DNA carries out PCR amplification, pcr amplification product is obtained；
B, Single base extension is carried out to the pcr amplification product with single strand dna shown in sequence 3, obtains extending production
Object；
C, the genotype or allele in the site rs1143634 in extension products described in Mass Spectrometer Method.
4th purpose of the invention is to provide any in product A-C.
It is any in product A-C provided by the invention, including IL-1 β gene in detection newborn or Fetal genome
The polymorphism or allele in the site rs1143634 or the substance of genotype；
A, prediction or auxiliary prediction fetus are the product that spontaneous pre-term easily sends out individual probability character；
B, the product of the risk of spontaneous pre-term occurs for assessment or aided assessment fetus to be measured；
C, identify or assist the product for the probability character for identifying that newborn is spontaneous pre-term.
And/or the polymorphism or genotype in the site IL-1 β gene rs1143634 in detection newborn's genome
Substance specifically includes shown in the primer pair and sequence 3 of the composition of single strand dna shown in single stranded DNA shown in sequence 1 and sequence 2
Single stranded DNA.
And/or the product is specially kit.
Above-mentioned product further includes being described below 1) -6) at least one of condition readable carrier:
1) it is that spontaneous pre-term is easy that the genotype in the site rs1143634 of IL-1 β gene, which is the fetus to be measured of CC genotype,
The individual probability of hair is less than or the candidate fetus to be measured for being less than CT genotype；
2) fetus to be measured that the allele in the site rs1143634 of IL-1 β gene is C is that spontaneous pre-term easily sends out individual
Probability is less than or the candidate allele that is less than is the fetus to be measured of T；
3) genotype in the site rs1143634 of IL-1 β gene is that spontaneous pre-term occurs for CC genotype fetus to be measured
Risk is less than or candidate is less than CT genotype fetus to be measured；
4) allele in the site rs1143634 of IL-1 β gene is that spontaneity occurs for the fetus to be measured of C allele early
The risk of production is less than or the candidate fetus to be measured for being less than T allele；
5) genotype in the site IL-1 β gene rs1143634 is that CC genotype waits for that detecting new-born baby is spontaneous pre-term
Probability is less than or the candidate CT genotype that is less than waits for detecting new-born baby；
6) it to detecting new-born baby is spontaneous pre-term that the allele in the site IL-1 β gene rs1143634, which is C allele,
The probability of youngster be less than or it is candidate be less than T allele to detecting new-born baby；
In above-mentioned application or above-mentioned product, fetus or newborn are Chinese.
The experiment proves that the present invention has studied the polymorphic site (G3953T (rs1143634)) of gene IL-1 β
A large amount of clinical and laboratory experiment has been carried out by the case-control study based on hospital with the correlation of premature labor neurological susceptibility
With the statistical analysis of large sample, distinguishes and carry the site IL-1 β G3953T T allele, at least one T allelotype (CT+
TT genotype) newborn occur SPTB (spontaneous pre-term) risk increase.The individual of CT genotype has premature labor higher
Neurological susceptibility.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1, the rs1143634 loci gene type for detecting IL-1 β gene
One, the design synthesis of the relevant primer of the rs1143634 loci gene type of IL-1 β gene is detected
It is to be measured using Sequenom company Genotyping Tools and MassARRAY Assay Design software design
The PCR amplification primer and Single base extension primer of SNP site, and biotech firm is transferred to synthesize.Detect IL-1 β gene
The site rs1143634 (the 14th on the 5th exon of IL-1 β gene, NG_008851.1:g.8967C > T；NM_000576.2:
C.315C > T NP_000567.1:p.Phe105=；The nucleotides sequence of IL-1 β gene is classified as sequence 1, and the site is in the sequence
3967；) genotype specific primer pair sequence are as follows: forward primer 5 '-
ACGTTGGATGAATCTCCCCAACATGAGTCC-3 ' (sequence 2 in sequence table)；Reverse primer 5 '-
ACGTTGGATGCAGGGATCTCCTGATCAGTT-3 ' (sequence 3 in sequence table).Extension for single base amplified reaction is drawn
Object sequence is 5 '-ATTGAGGGTGTAGACCTT-3 ' (sequence 4 in sequence table).
Two, the rs1143634 loci gene type of single base extension detection IL-1 β gene
1, the extraction of genomic DNA
Extract sample to be tested genomic DNA.
2, single base extension
1) PCR amplification
PCR amplification carries out in 384 orifice plates, and each reaction system total volume is 5 μ L, prepares PCR reaction system by table 1.
The component of each PCR reaction system of table 1
| Reagent | Volume (μ L) |
| 10 × PCR buffer | 0.5 |
| MgCl2(25mM) | 0.4 |
| dNTP mix(25mM) | 0.1 |
| HotStar Taq enzyme (5U/ μ L) | 0.1 |
| Ultrapure water | 1.9 |
| DNA shown in sequence 1 | 0.5 |
| DNA shown in sequence 2 | 0.5 |
| Total volume | 4 |
Take out above-mentioned 1 genome DNA sample prepared, adjustment injection volume is 1ul, in each 5ulPCR reaction system
Include template DNA 20-50ng, Hotstar Tag 0.5U, the 25mM dNTPs of every amplimer 0.5pmol, 0.1ul.
PCR response procedures are as follows: 94 DEG C 4 minutes；94 DEG C 20 seconds, 56 DEG C 30 seconds, 72 DEG C 1 minute, 45 circulation；72℃
3 minutes；4 DEG C of holdings.
Obtain PCR product.
2), PCR product alkaline phosphatase treatment:
PCR after reaction, by the above-mentioned PCR product 1) obtained SAP (shrimp alkaline
Phosphatase, shrimp alkaline phosphotase) processing, with remove system middle reaches from dNTPs, by table 2 preparation SAP reaction system.
2 SAP reaction system of table
Reaction condition are as follows: 37 DEG C 40 minutes；85 DEG C 5 minutes；4 DEG C of maintenances.
Obtain product after alkaline phosphatase treatment.
3) Single base extension
Single base extension is carried out after alkaline phosphatase treatment, 9 μ L of reaction system total volume is prepared by table 3
Single base extension system.
3 single base extension system of table
| Reagent | To each reaction, μ L |
| Water | 1.53 |
| 10 × single base extension buffer | 0.17 |
| Single base extension enzyme (1.7U/ul) | 0.3 |
| Product after alkaline phosphatase treatment | 7 |
| DNA molecular shown in sequence 3 | 1 |
| Total volume | 10 |
Reaction condition are as follows:
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℃
Obtain single base extension product.
3, purifying resin
By Clean Resin resin (Sequenom company, the U.S.) tiling into the resin plate of 6mg；Add 16 μ l water to above-mentioned
In the corresponding aperture of 2 obtained single base extension products；Resin after drying is poured into extension products plate, sealer, slow speed vertical
Rotation 30 minutes, comes into full contact with resin with reactant；Centrifugation makes resin sink to hole bottom, and the extension after obtaining purifying resin produces
Object.
4, Mass Spectrometer Method
Start MassARRAY Nanodispenser RS1000 point sample instrument (SEQUENOM company), above-mentioned 3 are obtained
Extension products after purifying resin move on 384 hole SpectroCHIP (Sequenom) chips (SEQUENOM company).By point sample
SpectroCHIP chip afterwards is analyzed using MALDI-TOF, and testing result uses 4.0 software of TYPER (sequenom) parting
And export result.As can be seen that the genotype in the site rs1143634 of IL-1 β gene is CC or TT or CT.
The research of embodiment 2, the polymorphic site rs1143634 genotype of IL-1 β gene and premature labor neurological susceptibility
Following research object is detected according to two method of embodiment 1:
1, the extraction of genomic DNA
Extract the genomic DNA of Chinese han population (727 prematures, 673 term infants) in vitro serum sample.
Above-mentioned in vitro serum from attached Aug. 1st children's hospital, General Hospital of Beijing Military Command 2 months in Mays, 2011 in 2009 and
The newborn that in March, 2014 accepts for medical treatment in September, 2014.All individuals are in the blood relationship for inhabit Beijing and surrounding area
Unrelated Chinese han population.
Case group is spontaneous pre-term (less than 37 pregnant weeks), and be divided into 3 groups according to pregnant week when birth: (33-36 is pregnant for middle premature labor
Week, 479)；Pole premature labor (< 32 pregnant weeks, 187)；Super premature labor (< 28 pregnant weeks, 61)；
Normal group (being equal to or more than 37 pregnant weeks) is the pregnant female life without premature rupture of fetal membranes and premature labor history randomly selected
673 term infants of single tire of production.
Newborn's selection criteria is that exclusion is slow with fetal anomaly, wound, Multiple organ diseases, preeclampsia, intrauterine growth
The newborn of the diseases such as premature labor caused by bleeding or other clinical diseases before slow, fetus crisis, childbirth.The prison of each participant
Shield signs informed consent form per capita, this research obtains batch of General Hospital of Beijing Military Command, P.L.A.'s Medical Ethics Committee
It is quasi-.
2, single base extension
Rs1143634 gene in the in vitro serum sample of each of above-mentioned 1 acquisition is detected according to two method of embodiment 1
Type.
Case group and the distribution of control group genotype meet Ha-temperature law of genetic equilibrium (P > 0.05), have good group
Body is representative, and TT genotype is free of in the genotyping result of case group.
The results are shown in Table 4, the genotype of the rs1143634 polymorphic site of the IL-1 β gene of case group and control group
With the distribution of gene frequency, in all samples, compared with the control group, T allele will be shown in the distribution frequency of case group
It writes and is higher than control group (χ 2=3.94, P=0.047, df=1), the distribution of CC, CT, TT genotype between case group and control group
Frequency has significant difference (χ 2=6.234, P=0.038, df=2).By Logistic regression analysis correct maternal age and
After sex of foetus, codominance (CT vs.CC is analyzed；TT vs.CC) hereditary pattern, it finds relative to CC genotype, CT genotype
(OR 1.69 related to the risk raising of SPTB occurs；95%CI, 1.06-2.69；P=0.038)；Analyze dominant (CT+TT
Vs.CC) hereditary pattern, discovery is for CC genotype, T allelotype (CT+TT genotype) and the risk liter that SPTB occurs
High correlation (OR 1.63；95%CI, 1.02-2.59；P=0.036)；Because genotyping result is free of TT genotype, recessive
(TT and CC+CT) hereditary pattern does not calculate；Superdominance (CT vs.CC+TT) hereditary pattern is analyzed, is found relative to CC+TT gene
Type, CT genotype (OR 1.69 related to the raising of the risk of SPTB；95%CI, 1.06-2.70；P=0.025)；With additivity mode
T equipotential dosage analysis is carried out, discovery T allele is incremented by (OR 1.56 uncorrelated to the risk raising of SPTB occurred；95%
CI, 0.99-2.45；P=0.053)；Illustrate the T allele of the rs1143634 polymorphic site of IL-1 β gene and occurs
The risk of SPTB increases correlation, is the risk allele that SPTB occurs；CT genotype is related to the risk raising of SPTB occurs,
It is the risk genotype that SPTB occurs.
The genotype and gene frequency of the rs1143634 polymorphic site of 4 case group of table and control group IL-1 β gene
Note: OR: odds ratio CI: confidence interval NA: not calculating
aOR value and P value are relatively got by CT genotype and CC+TT genotype
The above results show that phase occurs for the genotype in the site rs1143634 of IL-1 β gene or allele and premature labor
It closes, can be used to identify or assist to identify that detecting new-born baby be spontaneous pre-term probability character；It is also predicted that or assisting pre-
Surveying fetus to be measured is that spontaneous pre-term easily sends out individual probability character；It can also assess or spontaneity occurs for aided assessment fetus to be measured
The risk of premature labor；
It is above-mentioned identify or assist to identify include the following steps: to the method that detecting new-born baby is spontaneous pre-term probability character
The genotype for detecting the site rs1143634 of neonatal IL-1 β gene to be measured is CC or CT TT, CC genotype new life to be measured
Youngster is that spontaneous pre-term probability is less than or the candidate CT genotype that is less than waits for detecting new-born baby.
Or above-mentioned identify or assist to identify that the method that detecting new-born baby is spontaneous pre-term probability character include following step
Rapid: the allele in the site rs1143634 of detection neonatal IL-1 β gene to be measured is C or T, and C allele is to be measured new
Raw youngster is that spontaneous pre-term probability is less than or the candidate T allele that is less than waits for detecting new-born baby.
Above-mentioned prediction or auxiliary predict that fetus to be measured is that the method that spontaneous pre-term easily sends out individual probability includes the following steps:
The genotype for detecting the site rs1143634 of the IL-1 β gene of fetus to be measured is CC or CT, and the fetus to be measured of CC genotype is certainly
The premature labor of hair property easily send out individual probability and is less than or the candidate fetus to be measured less than CT genotype.
Or prediction or auxiliary predict that fetus to be measured is that the method that spontaneous pre-term easily sends out individual probability includes the following steps: to examine
The allele for surveying the site rs1143634 of the IL-1 β gene of fetus to be measured is C or T, and the fetus to be measured that allele is C is
Spontaneous pre-term easily sends out the fetus to be measured that individual probability is less than or candidate is T less than allele.
The method for the risk character that spontaneous pre-term occurs for above-mentioned assessment or aided assessment fetus to be measured includes the following steps:
The genotype for detecting the site rs1143634 of the IL-1 β gene of fetus to be measured is CC or CT, and CC genotype fetus to be measured occurs certainly
The risk of hair property premature labor is less than or candidate is less than CT genotype fetus to be measured.
Or the method for the risk character of spontaneous pre-term occurs for above-mentioned assessment or aided assessment fetus to be measured including walking as follows
Rapid: the allele for detecting the site rs1143634 of the IL-1 β gene of fetus to be measured is C or T, and allele is C tire to be measured
The risk that spontaneous pre-term occurs for youngster is less than or the candidate allele that is less than is the fetus to be measured of T.
When detecting fetus, the Cord blood of fetus or locating amniotic fluid can be taken to extract genomic DNA.
<110>hospital general, ground force, the Chinese People's Liberation Army
<120>IL-1 β gene pleiomorphism relevant to premature labor generation and its detection method
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 7020
<212> DNA
<213> Artificial Sequence
<220>
<223>
<400> 1
accaaacctc ttcgaggcac aaggcacaac aggctgctct gggattctct tcagccaatc 60
ttcattgctc aagtatgact ttaatcttcc ttacaactag gtgctaaggg agtctctctg 120
tctctctgcc tctttgtgtg tatgcatatt ctctctctct ctctctttct ttctctgtct 180
ctccctctcc ttccctctct gcctccctct ctcagctttt tgcaaaaatg ccaggtgtaa 240
tataatgctt atgactcggg aaatattctg ggaatggata ctgcttatct aacagctgac 300
accctaaagg ttagtgtcaa agcctctgct ccagctctcc tagccaatac attgctagtt 360
ggggtttggt ttagcaaatg cttttctcta gacccaaagg acttctcttt cacacattca 420
ttcatttact cagagatcat ttctttgcat gactgccatg cactggatgc tgagagaaat 480
cacacatgaa cgtagccgtc atggggaagt cactcatttt ctccttttta cacaggtgtc 540
tgaagcagcc atggcagaag tacctgagct cgccagtgaa atgatggctt attacaggtc 600
agtggagacg ctgagaccag taacatgagc aggtctcctc tttcaagagt agagtgttat 660
ctgtgcttgg agaccagatt tttcccctaa attgcctctt tcagtggcaa acagggtgcc 720
aagtaaatct gatttaaaga ctactttccc attacaagtc cctccagcct tgggacctgg 780
aggctatcca gatgtgttgt tgcaagggct tcctgcagag gcaaatgggg agaaaagact 840
ccaagcccac aatacaagga atccctttgc aaagtgtggc ttggagggag agggagagct 900
cagattttag ctgactctgc tgggctagag gttaggcctc aagatccaac agggagcacc 960
cagggtgccc acctgccagg cctagaatct gccttctgga ctgttctgcg catatcactg 1020
tgaaacttgc caggtgtttc aggcagcttt gagaggcagg ctgtttgcag tttcttatga 1080
acagtcaagt cttgtacaca gggaaggaaa aataaacctg tttagaagac ataattgaga 1140
catgtccctg tttttattac agtggcaatg aggatgactt gttctttgaa gctgatggcc 1200
ctaaacagat gaaggtaaga ctatgggttt aactcccaac ccaaggaagg gctctaacac 1260
agggaaagct caaagaaggg agttctgggc cactttgatg ccatggtatt ttgttttaga 1320
aagactttaa cctcttccag tgagacacag gctgcaccac ttgctgacct ggccacttgg 1380
tcatcatatc accacagtca ctcactaacg ttggtggtgg tggccacact tggtggtgac 1440
aggggaggag tagtgataat gtttcccatt tcatagtagg aagacaacca agtcttcaac 1500
ataaatttga ttatcctttt aagagatgga ttcagcctat gccaatcact tgagttaaac 1560
tctgaaacca agagatgatc ttgagaacta acatatgtct accccttttg agtagaatag 1620
ttttttgcta cctggggtga agcttataac aacaagacat agatgatata aacaaaaaga 1680
tgaattgaga cttgaaagaa aaccattcac ttgctgtttg accttgacaa gtcattttac 1740
ccgctttgga cctcatctga aaaataaagg gctgagctgg atgatctctg agattccagc 1800
atcctgcaac ctccagttct gaaatatttt cagttgtagc taagggcatt tgggcagcaa 1860
atggtcattt ttcagactca tccttacaaa gagccatgtt atattcctgc tgtcccttct 1920
gttttatatg atgctcagta gccttcctag gtggcccagc catcagccta gctaggtcag 1980
ttgtgcaggt tgggaggcag ccacttttct ctggctttat tttattccag tttgtgatag 2040
cctcccctag cctcataatc cagtcctcaa tcttgttaaa aacatatttc tttagaagtt 2100
ttaagactgg cataacttgt tggctgcagc tgtgggagga gcccattggc ttgtctgcct 2160
ggcctttgcc cccattgcct cttccagcag cttggctctg ctccaggcag gaaattctct 2220
cctgctcaac tttcttttgt gcacttacag gtctctttaa ctgtctttca agcctttgaa 2280
ccattatcat gccttaaggc aacctcagtg aagccttaat acggagcttc tctgaataag 2340
aggaaagtgg taacatttca caaaaagtac tctcacagga tttgcagaat gcctatgaga 2400
cagtgttatg aaaaaggaaa aaaaagaaca gtgtagaaaa attgaatact tgctgagtga 2460
gcataggtga atggaaaatg ttatggtcat ctgcatgaaa aagcaaatca tagtgtgaca 2520
gcattaggga tacaaaaaga tatagagaag gtatacatgt atggtgtagg tggggcatgt 2580
acaaaaaaga tgaacaaagt agaaatggga tttattctaa aagaatagcc tgtaaggtgt 2640
cagaaagccc acattctagt cttgagtctg cctctaacct gctgtgtgcc cttgagtaca 2700
cacttaacct ccttgagctt cagagaggga taatcttttt attttatttt attttatttt 2760
gttttgtttt gttttgtttt gttttatgag acagagtctc actctgttgc ccaggctgga 2820
gtgcagtggt acaatcttgg cttactgcat cctccacctc ctgagttcaa gcgattctcc 2880
ttcctcagtc tcctgaatag ctaggattac aggtgcaccc caccacaccc agctaatttt 2940
tgtattttta gtagagaagg ggtttcgcca tgttggccag gctggttttg aagtcctgac 3000
ctaaatgatt catccacctc ggcttcccaa agtgctggga ttacaggcat gagccaccac 3060
gcctggccca gagagggatg atctttagaa gctcgggatt ctttcaagcc ctttcctcct 3120
ctctgagctt tctactctct gatgtcaaag catggttcct ggcaggacca cctcaccagg 3180
ctccctccct cgctctctcc gcagtgctcc ttccaggacc tggacctctg ccctctggat 3240
ggcggcatcc agctacgaat ctccgaccac cactacagca agggcttcag gcaggccgcg 3300
tcagttgttg tggccatgga caagctgagg aagatgctgg ttccctgccc acagaccttc 3360
caggagaatg acctgagcac cttctttccc ttcatctttg aagaaggtag ttagccaaga 3420
gcaggcagta gatctccact tgtgtcctct tggaagtcat caagccccag ccaactcaat 3480
tcccccagag ccaaagccct ttaaaggtag aaggcccagc ggggagacaa aacaaagaag 3540
gctggaaacc aaagcaatca tctctttagt ggaaactatt cttaaagaag atcttgatgg 3600
ctactgacat ttgcaactcc ctcactcttt ctcaggggcc tttcacttac attgtcacca 3660
gaggttcgta acctccctgt gggctagtgt tatgaccatc accattttac ctaagtagct 3720
ctgttgctcg gccacagtga gcagtaatag acctgaagct ggaacccatg tctaatagtg 3780
tcaggtccag tgttcttagc caccccactc ccagcttcat ccctactggt gttgtcatca 3840
gactttgacc gtatatgctc aggtgtcctc caagaaatca aattttgccg cctcgcctca 3900
cgaggcctgc ccttctgatt ttatacctaa acaacatgtg ctccacattt cagaacctat 3960
cttcttcgac acatgggata acgaggctta tgtgcacgat gcacctgtac gatcactgaa 4020
ctgcacgctc cgggactcac agcaaaaaag cttggtgatg tctggtccat atgaactgaa 4080
agctctccac ctccagggac aggatatgga gcaacaaggt aaatggaaac atcctggttt 4140
ccctgcctgg cctcctggca gcttgctaat tctccatgtt ttaaacaaag tagaaagtta 4200
atttaaggca aatgatcaac acaagtgaaa aaaaatatta aaaaggaata tacaaacttt 4260
ggtcctagaa atggcacatt tgattgcact ggccagtgca tttgttaaca ggagtgtgac 4320
cctgagaaat tagacggctc aagcactccc aggaccatgt ccacccaagt ctcttgggca 4380
tagtgcaatg tcaattcttc cacaatatgg ggtcatttga tggacatggc ctaactgcct 4440
gtgggttctc tcttcctgtt gttgaggctg aaacaagagt gctggagcga taatgtgtcc 4500
atccccctcc ccagtcttcc ccccttgccc caacatccgt cccacccaat gccaggtggt 4560
tccttgtagg gaaattttac cgcccagcag gaacttatat ctctccgctg taacgggcaa 4620
aagtttcaag tgcggtgaac ccatcattag ctgtggtgat ctgcctggca tcgtgccaca 4680
gtagccaaag cctctgcaca ggagtgtggg caactaaggc tgctgacttt gaaggacagc 4740
ctcactcagg gggaagctat ttgctctcag ccaggccaag aaaatcctgt ttctttggaa 4800
tcgggtagta agagtgatcc cagggcctcc aattgacact gctgtgactg aggaagatca 4860
aaatgagtgt ctctctttgg agccactttc ccagctcagc ctctcctctc ccagtttctt 4920
cccatgggct actctctgtt cctgaaacag ttctggtgcc tgatttctgg cagaagtaca 4980
gcttcacctc tttcctttcc ttccacattg atcaagttgt tccgctcctg tggatgggca 5040
cattgccagc cagtgacaca atggcttcct tccttccttc cttcagcatt taaaatgtag 5100
accctctttc attctccgtt cctactgcta tgaggctctg agaaaccctc aggcctttga 5160
ggggaaaccc taaatcaaca aaatgaccct gctattgtct gtgagaagtc aagttatcct 5220
gtgtcttagg ccaaggaacc tcactgtggg ttcccacaga ggctaccaaa ttacatgtat 5280
cctactcatg gggcctaggg gttggggtga ccctgcactg ctgtgtccct aaccacaaga 5340
cccccttctt tcttcagtgg tgttctccat gtcctttgta caaggagaag aaagtaatga 5400
caaaatacct gtggccttgg gcctcaagga aaagaatctg tacctgtcct gcgtgttgaa 5460
agatgataag cccactctac agctggaggt aagtgaatgc tatggaatga agcccttctc 5520
agcctcctgc taccacttat tcccagacaa ccaccttctc cccgccccca tccctaggaa 5580
aagctgggaa caggtctatt tgacaatttt gcattaatgt aaataaattt aacataattt 5640
ttaactgcgt gcaaccttca atcctgctgc agaaaattaa atcattttgc cgatgttatt 5700
atgtcctacc atagttacaa ccccaacaga ttatatattg ttagggctgc tctcatttga 5760
tagacacctt gggaaataga tgacttaaag ggtcccatta tcatgtccac tccactccca 5820
aaattaccac cactatcacc tccagctttc tcagcaaaag cttcatttcc aagttgatgt 5880
cattctagga ccataaggaa aaatacaata aaaagcccct ggaaactagg tacttcaaga 5940
agctctagct taattttcac ccccccaaaa aaaaaaaatt ctcacctaca ttatgctcct 6000
cagcatttgg cactaagttt tagaaaagaa gaagggctct tttaataatc acacggaaag 6060
ttgggggccc agttacaact caggagtctg gctcctgatc atgtgacctg ctcgtcagtt 6120
tcctttctgg ccaacccaaa gaacatcttt cccatagcat ctttgtccct tgccccacaa 6180
aaattcttct ttctctttcg ctgcagagtg tagatcccaa aaattaccca aagaagaaga 6240
tggaaaagcg atttgtcttc aacaagatag aaatcaataa caagctggaa tttgagtctg 6300
cccagttccc caactggtac atcagcacct ctcaagcaga aaacatgccc gtcttcctgg 6360
gagggaccaa aggcggccag gatataactg acttcaccat gcaatttgtg tcttcctaaa 6420
gagagctgta cccagagagt cctgtgctga atgtggactc aatccctagg gctggcagaa 6480
agggaacaga aaggtttttg agtacggcta tagcctggac tttcctgttg tctacaccaa 6540
tgcccaactg cctgccttag ggtagtgcta agaggatctc ctgtccatca gccaggacag 6600
tcagctctct cctttcaggg ccaatcccca gcccttttgt tgagccaggc ctctctcacc 6660
tctcctactc acttaaagcc cgcctgacag aaaccacggc cacatttggt tctaagaaac 6720
cctctgtcat tcgctcccac attctgatga gcaaccgctt ccctatttat ttatttattt 6780
gtttgtttgt tttattcatt ggtctaattt attcaaaggg ggcaagaagt agcagtgtct 6840
gtaaaagagc ctagttttta atagctatgg aatcaattca atttggactg gtgtgctctc 6900
tttaaatcaa gtcctttaat taagactgaa aatatataag ctcagattat ttaaatggga 6960
atatttataa atgagcaaat atcatactgt tcaatggttc tgaaataaac ttcactgaag 7020
<210> 2
<211> 30
<212> DNA
<213> Artificial Sequence
<400> 2
acgttggatg aatctcccca acatgagtcc 30
<210> 3
<211> 30
<212> DNA
<213> Artificial Sequence
<400> 3
acgttggatg cagggatctc ctgatcagtt 30
<210> 4
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 4
attgagggtg tagacctt 18
Claims (1)
1. detecting in newborn's genomeIL-1βThe polymorphism in the site gene rs1143634 or the object of allele or genotype
Matter is identified or assists in preparation to identify newborn for the application in the probability character product of spontaneous pre-term；
The newborn is Chinese han population；
The genotype in the site rs1143634 is CC or CT；
The allele in the site rs1143634 is C or T；The site rs1143634 is located at human genome the 2nd dyeing
BodyIL-1βThe 14th on 5th exon of gene；
It is describedIL-1βThe nucleotides sequence of gene is classified as sequence 1.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517383A (en) * | 2011-12-12 | 2012-06-27 | 尤崇革 | Application of IL-6R gene SNP sites |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517383A (en) * | 2011-12-12 | 2012-06-27 | 尤崇革 | Application of IL-6R gene SNP sites |
Non-Patent Citations (3)
| Title |
|---|
| Identification of fetal and maternal single nucleotide polymorphisms in candidate genes that predispose to spontaneous preterm labor with intact membranes;Roberto Romero，et al;《Am J Obstet Gynecol》;20101231;第202卷;431.e1-34 |
| Interleukin-1 beta gene polymorphisms and preterm birth;Maximilian Schmid,et al;《European Journal 34 of Obstetrics & Gynecology and Reproductive Biology》;20121231;第165卷;33-36 |
| The Interleukin-1b +3953 Single Nucleotide Polymorphism:Cervical Protein Concentration and Preterm Delivery Risk;Rodney K. Edwards,et al;《American Journal of Reproductive Immunology》;20061231;第55卷;259–264 |
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