CN106337089A - LncRNA for diagnosing cerebral arterial thrombosis - Google Patents
LncRNA for diagnosing cerebral arterial thrombosis Download PDFInfo
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Abstract
The invention discloses lncRNA for diagnosing cerebral arterial thrombosis. The lncRNA is LOC105376505. According to the lncRNA for diagnosing cerebral arterial thrombosis, the LOC105376505 gene is related to occurrence and development of cerebral arterial thrombosis, expression in cerebral arterial thrombosis patients is reduced, and researching of pathogenesis of cerebral arterial thrombosis is further enriched.
Description
Technical field
The invention belongs to biomedicine field, it is related to a kind of lncrna for cerebral arterial thrombosis diagnosis, specifically should
Lncrna is loc105376505.
Background technology
Cerebrovascular disease is one of three big diseases of serious harm human health together with cardiovascular disease and tumour, and it has height
The feature of the incidence of disease, high disability rate and high mortality, is also the big cause of death of the mankind three together with cardiovascular disease and tumour simultaneously
One of.Cerebrovascular disease leads to the nervous system disease of local nerve afunction mainly due to brain tissue blood circulation disorder
Disease, can be divided into ACVD and chronic cerebrovascular disease.Acute cerebrovascular diseases is commonly referred to as cerebral apoplexy again, in the U.S., often
Year, about 800,000 people suffered from cerebral apoplexy class disease, and the incidence of disease increases with the increase at age, and this gives the society of its continuous astogeny
Serious problem can be brought.In China, Cerebral Vascular Disease rate rises year by year, has occupied population lethal and disable the first of reason
Position.And the fast development with economic level, constantly the increasing of China human mortality Aging Problem, and unsound life style
Deng many factors so that the ischemic cerebrovascular disease incidence of disease increases year by year.Cerebral apoplexy can be divided into cerebral arterial thrombosis again
(ischemic stroke, is) and hemorrhagic apoplexy (hemorrhagic stroke).Cerebral arterial thrombosis accounts for whole brain soldiers
The 80% about of the middle incidence of disease, and its incidence of disease increases with the growth at age.The generation of cerebral arterial thrombosis mainly by
In the formation of cerebral embolism and local thrombus, this is probably the blood vessel being caused by atherosclerotic (atherosclerosis, as)
Narrow, thromboembolism, or cause cerebral embolism with blood flow to brain from the embolus of heart；And the blood that a variety of causes causes
Pipe damages, vascular inflammation is also one of major reason.Many reasons cause brain blood supply obstacle, and cause injured cerebral tissue
Irreversibility damage so that brain tissue ischemia, anoxic lead to final necrosis, cause a series of neurologic impairment to patient
And obstacle.
Cerebral arterial thrombosis is inherent cause and environmental factor coefficient multiple-factor complex disease, except environmental factor
Impact beyond, inherent cause plays a very important role in the pathogenic process of cerebral apoplexy, existing many molecular genetics
Research also show cerebral apoplexy and has very high genetic predisposition.Inherent cause may be by Traditional Factors are lured
Send out, or as independent risk factor, eventually affect histoorgan.Depending on brain ct the diagnosis of cerebral arterial thrombosis at present more
How scanning, brain mri inspection, dsa, mra, TCD inspection etc., diagnose to cerebral arterial thrombosis earlier
And prevention, it is the important topic of current research.
Recently it has been found that except there being microrna, this has robust adaptive control and epigenetic is repaiied in human genome
The presence of small non-coding rna of decorations function, also there are ten hundreds of another kind of non-codings rna is long-chain non-coding rna
(long non-coding rna, lncrna) long-chain non-coding is the rna molecule that a class transcript length is more than 200nt, they
There is no the function of encoding proteins.Lncrna is initially believed to be the accessory substance of rna polymerase ii transcription, is subgenomic transcription
" noise, rubbish ", does not have biological function.However, it has recently been demonstrated that they can be methylated by dna, histone
Multiple different mechanism such as modification, post-transcriptional control, rna interference, imprinted genes, in multiple aspects affect gene expression water
Flat.Therefore, lncrna is likely to become the new direction of cerebral arterial thrombosis research.It has been reported that microrna is lacking in prior art
There is to have important function in development and resistance in courageous and upright cerebral apoplexy, but the same lncrna as non-coding product, its life
Thing function and its effect in cerebral arterial thrombosis generation evolution are unclear, require further study.
At present, the lncrna of the unconventionality expression finding in illing tissue can relate to each system of whole body, and distribution is more wide
General, but due to lncrna substantial amounts, still it is in the starting stage currently for research in cerebral apoplexy field for the lncrna, because
The molecular basis that this seeks lncrna cerebral arterial thrombosis is significant.
Content of the invention
In order to make up the deficiencies in the prior art, an object of the present invention, provide a kind of detection long-chain non-coding rna
The product of loc105376505, the early diagnosis for realizing cerebral arterial thrombosis provides basis.
The second object of the present invention, provides a kind of diagnostic method of cerebral arterial thrombosis, by detecting lncrna
Whether the expression of loc105376505 suffers from cerebral arterial thrombosis diagnosing patient.
The third object of the present invention, provides a kind of molecular marker, and the index as disease detection is applied to clinic.
The fourth object of the present invention, provides a kind of development of the generation with cerebral arterial thrombosis closely related gene, is scarce
The scientific research of courageous and upright cerebral apoplexy provides theoretical foundation.
To achieve these goals, the present invention adopts the following technical scheme that
The invention provides long-chain non-coding rna loc105376505 is in the product preparing diagnosing ischemia cerebral apoplexy
Application.Wherein, loc105376505 refers to gene or the mrna from this genetic transcription, in the mankind, short positioned at No. 11 chromosomes
Arm the 1st area the 5th carries on the 5th subzone.
Further, the sequence of described loc105376505 is as shown in seq id no.1.
Further, described loc105376505 expresses downward in ischemic cerebral stroke patients.
Further, described product includes: by sequencing technologies, nucleic acid hybridization technique, nucleic acid amplification technologies or immunoassays
Method detect loc105376505 gene expression.
Further, described nucleic acid amplification technologies are selected from PCR (pcr), reverse transcriptase polymerase chain reaction
(rt-pcr), the amplification (tma) of transcriptive intermediate, ligase chain reaction (lcr), strand displacement amplification (sda) and be based on nucleic acid sequence
The amplification (nasba) of row.Wherein, pcr needs, before amplification, rna reverse transcription is become dna (rt-pcr), tma and nasba directly expands
Increase rna.
Generally, pcr uses the annealing of denaturation, primer pair and opposite strand and multiple circulations of primer extend, with index side
Formula increases the copy number of target nucleic acid sequence；Reverse transcriptase (rt) is then used for from the complementary dna (cdna) of mrna preparation by rt-pcr,
Then cdna is expanded the multiple copies producing dna by pcr；Tma is in the temperature of substantial constant, ionic strength and ph
Under the conditions of autocatalytically synthesize multiple copies of target nucleic acid sequence, multiple rna copies of wherein target sequence are autocatalytically given birth to
Become other copy, tma optionally includes using blocking, partly, dwell section and other modify part, to improve tma process
Sensitivity and the degree of accuracy；Lcr is using two groups of complementary dna oligonucleotides with the adjacent area hybridization of target nucleic acid.Dna few nucleosides
Acid thermal denaturation, hybridization and connection repeat multiple circulation in be covalently attached by dna ligase, to produce detectable double-strand
Connect oligonucleotide product；Sda is using multiple circulations of following steps: primer sequence pair is moved back with the opposite strand of target sequence
Fire, carries out primer extend to produce (hemiphosphorothioated) of double-strand half thiophosphorylation under there is dntp α s
Primer extension product, the nicking of endonuclease that semi-modified restriction enzyme enzyme recognition site is carried out mediation, and from cutting
The polymerase-mediated thing drawing that mouthful 3' end is carried out extends to be put for next round primer annealing, nicking and chain with replacing existing chain producing
The chain changing, thus cause the geometry of product to expand.
The invention provides in a kind of vitro detection sample above-mentioned lncrna loc105376505 expression product,
Described product includes preparation, chip or kit.Described " sample " include cell, tissue, internal organs, cerebrospinal fluid, body fluid (blood,
Lymph liquid etc.), digestive juice, expectoration, alveole bronchus cleaning fluid, urine, excrement etc..Preferably, described sample is tissue, blood.
In the specific embodiment of the present invention, selected sample is blood.
Further, described preparation, chip or kit include specific primer for loc105376505 to or probe.
Further, the sequence of described specific primer pair is as shown in seq id no.2 and seq id no.3.
Further, described specific primer is visited to for sybr green, taqman probe, molecular beacon, double cross
Pin, the detection of combined probe.
The invention provides a kind of application in the instrument preparing diagnosing ischemia cerebral apoplexy for product recited above.
In the present invention, " probe " refers to the molecule being combined with the particular sequence of another molecule or subsequence or other parts.Remove
Non- indicate otherwise, term " probe " be often referred to can by complementary base pairing with another polynucleotides (often referred to as " many nucleosides of target
Acid ") polynucleotide probes that combine.According to the preciseness of hybridization conditions, probe energy and to lack sufficient sequence complementary with this probe
Property target polynucleotide combine.Probe can make direct or indirect mark, and its scope includes primer.Crossing system, including, but not
It is limited to: solution, solid phase, mixed phase or in situ hybridization determination method.
Described probe has the base sequence with the specific base sequence complementary of target gene.Here, so-called " complementary ",
As long as hybridizing, can not be complete complementary.These polynucleotides are commonly angled relative to this specific base sequence to be had
More than 80%, preferably more than 90%, more preferably more than 95%, particularly preferred 100% homology.These probes can be dna,
Can also be rna, furthermore it is possible to be to pass through pna (polyamide nucleic in one part or whole nucleotide
Acid, peptide nucleic acid), lna (registration mark, locked nucleic acid, bridged nucleic acid, Cross-linked core
Acid), ena (registration mark, 2 '-o, 4 '-c-ethylene-bridged nucleic acids), gna (glycerol
Nucleic acid, glycerine nucleic acid), the artificial replacement nucleic acid such as tna (threose nucleic acid, threose nucleic acid) obtains
Polynucleotides.
Term " homology " refers to the degree of complementarity.There may be Homoeology or complete homology is (that is, same
Property).The sequence of partial complementarity is that the nucleic acid molecules of suppression complete complementary at least in part are miscellaneous with " substantially homology " target nucleic acid
The nucleic acid molecules handed over.Fully-complementary sequence can pass through to survey using hybridization under low stringency with the suppression of the hybridization of target sequence
Fixed (southern or northern trace, solution hybridization etc.) is checked.Substantially the sequence of homology or probe will compete simultaneously
Suppress nucleic acid molecules combination (that is, hybridizing) with target under low stringency of complete homology.This is not to say, low strict
Property condition be so that allow non-specific binding condition；Low stringency require two sequences each other be combined into spy
The interaction of the opposite sex (i.e., selectively).Not existing of non-specific binding can be (for example, low by using substantially incomplementarity
In about 30% homogeneity) the second target tested；In the case of there is not non-specific binding, probe will not be with second
Incomplementarity target hybridization.
Term " hybridization " in the present invention is used for referring to the pairing of complementary nucleic acid.Hybridization and intensity for hybridization are (that is, between nucleic acid
Association intensity) affected by such as following factor: the complementarity between nucleic acid, the stringency of involved condition, formation
The tm and nucleic acid of crossbred in g:c ratio.The individual molecule being contained within the pairing of complementary nucleic acid in its structure is referred to as " self
Hybridization ".
In the present invention, gene detecting kit or genetic chip be can be used for detection including loc105376505 gene
The expression of multiple genes (for example, the multiple genes related to cerebral arterial thrombosis), by multiple marks of cerebral arterial thrombosis
Will thing is detected simultaneously, is greatly improved the accuracy rate of cerebral arterial thrombosis diagnosis.
It would be recognized by those skilled in the art that the practicality of the present invention is not limited to the marker gene to the present invention
The gene expression of any specific variants carries out quantitation.As nonrestrictive example, marker gene can have seq id no.1
The cdna sequence specified.In some embodiments, it has the same or analogous cdna sequence with listed sequence at least 85%
Row, such as above-mentioned listed sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least
99% same or analogous cdna sequence.
Nucleic acid hybridization technique in the present invention include but is not limited in situ hybridization (ish), microarray and southern or
Northern trace.In situ hybridization (ish) be a kind of be used mark complementary dna or rna chain as probe with position tissue one
Part or section (in situ) or if tissue is sufficiently small, for the specific dna in whole tissue (full organization embedding ish) or
The hybridization of rna sequence.Dna ish can be used for determining the structure of chromosome.Rna ish is used for measurement and position tissue section or complete
Mrna in organization embedding and other transcripts (for example, ncrna).Generally sample cell and tissue are processed solid with original position
Targeting transcript, and increase the entrance of probe.Probe is hybridized with target sequence at high temperature, then washes unnecessary probe off.Point
Not using autoradiograph, fluorescence microscopy or immunohistochemistry, to the base with radiation, fluorescence or antigenic mark in tissue
The probe of mark is positioned and quantitative.Ish also can pass through radioactivity or other nonradioactive labelings using two or more
The probe of substance markers, to detect two or more transcripts simultaneously.
Term " microarray ", including but not limited to: dna microarray (for example, cdna microarray and oligonucleotide microarray),
Protein microarray, micro-array tissue, transfection or cell microarray, chemical compound microarray and Antibody microarray.Commonly referred to as
For genetic chip, dna chip or biochip dna microarray be microcosmic dna point set, these point be connected to the surface of solids
On (for example, glass, plastics or silicon), formed for several thousands genes are carried out with expression pattern analysis or expression prison simultaneously
The array surveyed.Fixing dna fragment is referred to as probe, and it is thousands of to can be used in single dna microarray.Microarray can be used for passing through
Compare the gene expression in disease and normal cell and identify disease gene or transcript (for example, ncrna).Microarray can use
Multiple technologies are manufactured, including but not limited to: be printed onto with apicule needle on slide, carried out using prefabricated mask photoetching,
Carry out the electrochemical method on photoetching, ink jet printing or microelectrode array using dynamic micro mirror element.
Southern and northern trace is respectively used to detect specific dna or rna sequence.Make to extract from sample
Dna or rna fracture, separated by electrophoresis on matrix gel, be then transferred on molecular filter.Make filter combine dna or
Rna with and the complementary label probe of sequence of interest hybridize.Detection is attached to the hybridization probe of filter.A kind of change of this program
Change form is reverse northern blot, and the substrate nucleic acid being wherein fixed to film is the set of detached dna fragment, and probe is
From tissue extraction and the rna that marked.
Term " diagnosis " in the present invention refers to the S＆S by disease or genetic analysis, pathological analysis, tissue
The identification disease such as credit analysis.
Advantages of the present invention and beneficial effect:
Present invention finds a kind of molecular marker of diagnosing ischemia cerebral apoplexy, by detecting mark
The expression of loc105376505 can judge to early stage cerebral arterial thrombosis, thus carrying out early intervention, improving and suffering from
The survival rate of person.
The invention provides a kind of closely related gene of generation development with cerebral arterial thrombosis, it is cerebral arterial thrombosis
Scientific research provide theoretical foundation.
Brief description
Fig. 1 shows the difference expression gene using cDNA microarray ischemic cerebral stroke patients；
Fig. 2 shows using qpcr detection expression in ischemic cerebral stroke patients for the loc105376505.
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 restriction 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) condition described in, or according to the condition proposed by manufacturer.
The embodiment 1 screening gene marker related to cerebral arterial thrombosis
1st, sample collection
Respectively collect the blood of 10 normal human bloods and ischemic cerebral stroke patients, the equal informed consent of patient, above-mentioned own
The acquirement of sample is all by the agreement of the committee of organizational ethics.
2nd, the preparation of rna sample
1) 12000rpm high speed centrifugation 10min after clinical serum sample collection, is repeated 2 times, -80 DEG C of guarantors of gained serum sample
Deposit；
2) frozen 4 DEG C of defrostings of serum sample；
3) serum sample drawing 250 μ 1 is managed to 1.5m1ep, plus 750 μ 1trizol ls reagent, and piping and druming mixes, quiet
Put 5min；
4) (chcl imitated by chlorination3: trizol 250 μ 1:750 μ 1), overturn and mix, stand 15min；
5) 4 DEG C, 12000rpm, it is centrifuged 15min；
6) careful supernatant liquid of drawing is to a new ep pipe；
7) add appropriate isopropanol (isopropanol: trizo1 500 μ 1:750 μ 1), overturn and mix, stand 10min；
8) 4 DEG C, 12000rpm is centrifuged 10min；
9) abandon upper strata centrifugate, plus 75% ethanol (rna enzyme-deactivating)；
10) 4 DEG C, 8000rpm is centrifuged 5min；
11) retain precipitation, abandon upper strata centrifugate, room temperature stands 5-10min；
12) appropriate depc water dissolves precipitation, takes appropriate rna solution to survey concentration and observe rna extracting situation；
13) mark title, concentration and the date, -80 DEG C of preservations.
3rd, reverse transcription and mark
With low rna input linear amplification kit, mrna reverse transcription is become cdna, use cy3 simultaneously
Labelling experiment group and control group respectively.
4th, hybridize
Genetic chip adopts Kang Cheng biology-human lncrna array, carries out miscellaneous by the step of chip operation instructions
Hand over.
5th, data processing
Chip agilent scanner scanning after hybridization, resolution ratio is 5 μm, and scanner is automatically with 100% and 10%pmt
Respectively scanning 1 time, 2 times result agilent software merges automatically.Scan image data using feature extraction at
Reason analysis, the initial data application bioconductor program bag obtaining carries out follow-up data process.Last ratio value is experiment
Group and control group.Differential gene screening criteria: ratio >=4 are up-regulated gene, ratio≤0.25 is down-regulated gene.
6th, result
Result as shown in figure 1, compared with healthy human blood, expression in ischemic cerebral stroke patients for the loc105376505
Level is substantially less than the expression of Healthy People.
The differential expression of embodiment 2 qpcr sequence verification loc105376505 gene
1st, loc105376505 gene differential expression is carried out with large sample qpcr checking.Receive according to the sample in embodiment 1
Mode set selects each 100 of the blood sample sample of normal person and ischemic cerebral stroke patients.
2nd, rna extraction step is with embodiment 1.
3rd, reverse transcription:
(1) reverse transcription reaction:
Using 25 μ l reaction systems, each sample takes the total rna of 1 μ g as template rna, is separately added into following in pcr pipe
Component: depc water, 5 × RT Buffer, 10mm dntp, 0.1mm dtt, 30 μm of oligo dt, 200u/ μ l m-mlv,
Template rna.42 DEG C of incubation 1h, 72 DEG C of 10min, of short duration centrifugation.
(2) design of primers:
The primer sequence of loc105376505 gene is:
Forward primer: 5 '-aactgaagaaccacaagagaag-3 ' (seq id no.2)
Reverse primer: 5 '-ttgctgtccaggagaagg-3 ' (seq id no.3)
The primer sequence of house-keeping gene gapdh is:
Forward primer: 5 '-ccgggaaactgtggcgtgatgg-3 ' (seq id no.4)
Reverse primer: 5 '-aggtggaggagtgggtgtcgctgtt-3 ' (seq id no.5)
(3) qpcr amplification inspection:
Using 25 μ l reaction systems, each sample arranges 3 parallel pipes, and all amplified reactions are all in triplicate above to protect
The reliability of card result.
Prepare following reaction system: sybr green PCR system 12.5 μ l, forward and reverse primer (5 μm) is each
1 μ l, template cdna 2.0 μ l, no enzyme water 8.5 μ l.Operations are all carried out on ice.
Amplification program is: 95 DEG C of 60s, (95 DEG C of 15s, 60 DEG C of 15s, 72 DEG C of 45s) × 40 circulations.
Using sybr green as fluorescent marker, light cycler fluorescence real-time quantitative pcr instrument carries out pcr anti-
Should, purpose band is determined by melt curve analysis analysis and electrophoresis, δ δ ct method carries out relative quantification.
3rd, statistical method
Experiment all to complete according to being repeated 3 times, and result data is all to be represented in the way of mean+SD,
Statistical analysis is carried out using spss18.0 statistical software, difference between the two adopts t inspection it is believed that when p < has when 0.05
Statistically significant.
4th, result
Result as shown in Fig. 2 compared with Healthy People, under loc105376505 gene is expressed in ischemic cerebral stroke patients
Adjust, difference has statistical significance (p < 0.05), consistent with rna-sep result.
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 improve and modify also by the protection domain falling into the claims in the present invention.
sequence listing
<110>The First Hospital of Medical College of Shantou University
<120>a kind of lncrna for cerebral arterial thrombosis diagnosis
<160> 5
<170> patentin version 3.5
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ctcctgtctc agcctcccga gtagctggaa ctcccaagct cagagtggag aatgaggttg 60
catctcagga tggccacagc gccttccttc ccaagccctt tcttcagcag ccgtggtggg 120
agctgcagcc gaaaagcccc tttggagttg cagctagagg gtgccttcca ggagctgctg 180
ggccagcttc accaatgcct gcagacggtc aggacagatt cagagaagct ctgggtcaga 240
acgagggtga aaagcccctt cctcgagctt ccggtcgaac agccccatgg gggtgccctg 300
accttcttgc tctaatgcag ctgctgttcc ccggggcagc gactcccctt ctctccctcc 360
caagtgcggc cctgccctct tcacaggcac acaccccaca gcacgtgcag tgcctcaagg 420
cgcccaaggt gcacggggag gcccatcagt gctgttcccc aagcctccag ctttcttgct 480
ggaatctttc tttcccaccc ctttgcagct tggcatgagc ttgtcacttg aggtggccag 540
tgagatgttg tgggcatgag aggtggtgcc aagagatcag gcgtgactcc acctcccgtt 600
cccacctcgg aggcctggca gcacacgctg ggggctcccc aacctgggcg gctgggcgag 660
gacgaggcgc agaccccaca cctgccacag gcccacactg gagggagtca gcggctgagg 720
gtcgggccgt tcgttaccac cgcacagcct ggctcatcct ggctgacttg gggcagtgct 780
gcccaggcca ggcggagcag ggaggacgtg gtggggtgtt tggaaggcgg agcaagaacc 840
taataaatgt gagccctgga ctctgaggcc gggcagcgaa tgttctgcgg gtgctgtctg 900
cggccggatg tgggttttgg catcatgagc tcaggagaga actggccggt gggtgagcag 960
gtgcgaaagg aaaccgagaa aacccggggg gtactttgtg ggttaaaaga ggcaactgcc 1020
tttccatcct gaccagcaga agggaagact gcagagaccc tcgaatgtcg atctcagcgg 1080
ttgcggctag gatggaatca agagcaggcc accgcaccca ctgctgacaa atctccagcc 1140
tgggtgacag agccagaccc tgctgtcagg cctccgagcc caagcctgga cgtgtacatc 1200
cagatggcct gaggcaactg aagaaccaca agagaagtga aaatggccag ttcctgcctt 1260
aactgacgac attgcctcat gaaattcctt ctcctggaca gcaagaaagg agctgaggaa 1320
gccactcaac tctccagcac ccaccctgtg cggccacgga ggaccgtggg gatgggaagt 1380
gtcccctcca ccgaggacag ccctcatctc ttcccggaag ttgctccccg gcgtcagcgt 1440
cctcacccag ccacccctgg gactttggag ttgctgcaga gcatgactgc tgggatctgc 1500
cttcttccct tccaaaccgt cgaagcccct gtccctgctc cttggttgaa tactgggggc 1560
gtgggcagaa aactcttcat ctttttaatt cagggccctc cagcttggga ggagacacag 1620
ctaggctgga tgcggcaact tgtgtacttt tgaccttcat gtagctggag gacaccgggt 1680
tgcccttgag gaagcaggtg tattttgcct gcaggaaggg cagtcttctg agtacctcgc 1740
aggtgtctgg atgtggtgcg cattagtgct gttcatcaga caccttcagc tgtcctctgg 1800
gcagatggca gggttgtgtc ttcctgtgga gttccttcct ttcgcctcag tgaccacgga 1860
gacgcatgtc gaggtgtgac ctcgcctggg cctctgcatg agggtgggga gcagagtcct 1920
ccttagtggg agtggagcgt gcacgggacg taattacgtg gggccatttg ttacgcagcc 1980
taacacagcc tgtcctggct gatgcagatg gccattgcag cagtcatctg caggtcctca 2040
gcttagtctc accctcactg gagatcttag ccttggcctc acaggtgctc cagacctttt 2100
cctggcagaa ttttggtgac ttcagaaata tgatgactgc tgaacccact tctcccagca 2160
tttctatttg tggttgcctg gtgtatttaa agtctgtgtt gatgggtgca tatattttta 2220
acgtttccct attgtgaaat ttaataaatt tacagcaaaa tgtataaaat acgaaagtaa 2280
aaa 2283
<210> 2
<211> 22
<212> dna
<213>artificial sequence
<400> 2
aactgaagaa ccacaagaga ag 22
<210> 3
<211> 18
<212> dna
<213>artificial sequence
<400> 3
ttgctgtcca ggagaagg 18
<210> 4
<211> 22
<212> dna
<213>artificial sequence
<400> 4
ccgggaaact gtggcgtgat gg 22
<210> 5
<211> 25
<212> dna
<213>artificial sequence
<400> 5
aggtggagga gtgggtgtcg ctgtt 25
Claims (10)
1. application in the product preparing diagnosing ischemia cerebral apoplexy for the long-chain non-coding rna loc105376505.
2. application according to claim 1 is it is characterised in that the sequence such as seq id no.1 of described loc105376505
Shown.
3. application according to claim 1 and 2 is it is characterised in that described loc105376505 suffers from cerebral arterial thrombosis
In person, expression is lowered.
4. application according to claim 1 and 2 is it is characterised in that described product includes: miscellaneous by sequencing technologies, nucleic acid
The method of friendship technology, nucleic acid amplification technologies or immunoassays detects the expression of loc105376505 gene.
5. according to claim 4 application it is characterised in that described nucleic acid amplification technologies be selected from PCR,
Reverse transcriptase polymerase chain reaction, the amplification of transcriptive intermediate, ligase chain reaction, strand displacement amplification and based on nucleotide sequence
Amplification.
6. the product of the lncrna loc105376505 expression described in claim 1 or 2 in a kind of vitro detection sample
Product are it is characterised in that described product includes preparation, chip or kit.
7. product according to claim 6 it is characterised in that it is characterized in that, described preparation, chip or kit include
For loc105376505 specific primer to or probe.
8. product according to claim 7 is it is characterised in that the sequence such as seq id no.2 of described specific primer pair
With shown in seq id no.3.
9. the product according to claim 7 or 8 it is characterised in that described specific primer to for sybr green,
Taqman probe, molecular beacon, double cross probe, the detection of combined probe.
10. application in the instrument preparing diagnosing ischemia cerebral apoplexy for the product described in any one of claim 6-9.
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| CN201611053225.9A CN106337089B (en) | 2016-11-24 | 2016-11-24 | A kind of lncRNA for cerebral arterial thrombosis diagnosis |
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| CN201611053225.9A CN106337089B (en) | 2016-11-24 | 2016-11-24 | A kind of lncRNA for cerebral arterial thrombosis diagnosis |
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| CN106337089B CN106337089B (en) | 2018-08-21 |
Family
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