CN110396542A - A kind of LncRNA marker and its application in diabetes - Google Patents
A kind of LncRNA marker and its application in diabetes Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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- 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
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- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
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- C—CHEMISTRY; METALLURGY
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- C12Q2600/158—Expression markers
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- C—CHEMISTRY; METALLURGY
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- 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
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Abstract
The invention belongs to field of biological detection, and in particular to a kind of LncRNA marker and its application in diabetes.The present invention successfully detects that LncRNA molecular marked compound NONHSAT024449 has obvious differential expression in diabetic's blood, compared with normal population control group, diabetic population experimental group patient has the LncRNA molecular marked compound NONHSAT024449 of low expression, based on this discovery, LncRNA molecular marked compound NONHSAT024449 can be used as diabetes molecular marker or target spot is applied to diabetes clinical diagnosis or targeted therapy, by detecting the specific primer of LncRNA molecular marked compound NONHSAT024449 provided by the present invention to the early screening that diabetes may be implemented, prevention/assessment diabetes occurrence risk, by LncRNA molecular marked compound NONHSAT02 4449 are applied to prepare diabetes diagnosis product or prepare in diabetes mellitus prevention/assessment product, are conducive to further elucidate diabetes occurrence and development mechanism, signal path etc., great application prospect and theoretical value.
Description
Technical field
The invention belongs to field of biological detection, and in particular to a kind of LncRNA marker and its application in diabetes.
Background technique
Diabetes are a kind of multi-pathogenesis metabolic diseases characterized by hyperglycemia, can lead to various tissues, especially eye,
Kidney, heart, blood vessel, the chronic lesion of nerve and dysfunction.The occurrence and development of diabetes are a chronic processes, early
The symptom of phase is not obvious.Existing research shows that long-chain non-coding RNA (LncRNA) can by regulation mRNA transcription and translation or
It modifies to have the function that influence Occurrence and development of disease.PCR has the specificity and sensibility of height, can be by the change of very little
Change is reflected by intuitive numerical value.Have the research that related gene or the detection of LncRNA are carried out by means such as PCR,
But it is concentrated mainly on the nervous system disease, in the diseases such as cardiovascular disease and tumor and cancer.
Currently, the researchs such as differential expression and diagnostic value, mechanism of action are not at home and abroad in diabetic population by LncRNA
It appears in the newspapers, research of the LncRNA marker in diabetes regulatory mechanism is still in infancy.Therefore, by analyzing, studying
LncRNA plays a significant role early screening in the occurrence and development mechanism of diabetes, and controls for the gene target of diabetes
Treating offer may.
Summary of the invention
In order to make up some shortcomings of the prior art, an object of the present invention, provide one kind has spy in diabetes
The LncRNA marker of opposite sex expression provides the specific primer pair for expanding the LncRNA marker, and applies primer pair
It carries out diabetes early screening aided assessment or forecast assessment information evaluation is collected.
The second object of the present invention, provides assessment or the diagnostic products of a kind of diabetes, which examines for diabetes
Break sensitivity and specificity with higher.
To achieve the goals above, the present invention adopts the following technical scheme:
The present invention provides a kind of LncRNA marker, the LncRNA marker is NONHSAT024449, the LncRNA mark
Remember object low expression in diabetic.
Further, the nucleotide sequence of the LncRNA marker is as shown in SEQ ID NO.1.
Further, the specific primer pair of the LncRNA marker, upstream primer nucleotide sequences such as SEQ ID
Shown in NO.2；Primer nucleotide sequences are as shown in SEQ ID NO.3 downstream.It is tested by largely screening property, according to LncRNA
The PCR primer sequence of marker carries out PCR amplification to diabetic's blood sample, and specific fragment can be obtained at 271bp
Purpose band, amplified production determine the expression quantity of LncRNA in sample to be detected by purpose band.
The present invention also provides application of the above-mentioned LncRNA marker in preparation assessment or diagnosis diabetic Products.Into
One step, the product include: to be examined by RT-PCR, real-time quantitative PCR, in situ hybridization, chip or high-flux sequence platform
The expression of LncRNA marker NONHSAT024449 is surveyed to assess or diagnose diabetes.Wherein, described to be diagnosed with RT-PCR
The product of diabetes includes at least the primer of a pair of of specific amplified NONHSAT024449 gene；It is described to be diagnosed with real-time quantitative PCR
The product of diabetes includes at least the primer of a pair of of specific amplified NONHSAT024449 gene；It is described to diagnose sugar in situ hybridization
The product of urine disease includes: the probe with the nucleic acid array hybridizing of NONHSAT024449 gene；It is described to diagnose diabetes with chip
Product includes the probe with the nucleic acid array hybridizing of NONHSAT024449 gene.
The present invention provides a kind of product, the product can measure above-mentioned LncRNA marker NONHSAT024449 in sample
Content, wherein NONHSAT024449 expresses downward in diabetic." sample " described in invention includes cell, group
It knits, internal organs, body fluid (blood, lymph etc.), digestive juice, expectoration, alveole bronchus cleaning solution, urine, excrement etc..Preferably, institute
Stating sample is blood.The products application is in the tool of preparation assessment or diagnosis diabetes.
Further, the product includes kit, probe, chip or preparation.
Further, the kit includes the specific primer pair of above-mentioned LncRNA marker.The kit further includes
DNTP, random primer, reducing agent, RNase inhibitor, reverse transcriptase, MgCl2With one of PCR buffer etc. or a variety of
Combination, in addition, the kit can also contain standard items and/or reference substance.
Further, mentioned reagent box includes also preferably some other auxiliary reagent, and the auxiliary reagent is quantitative pcr amplification
Conventional use of some reagents in kit, the characteristic of these reagents and their preparation method are those skilled in the art
Known；The reagent such as (but not limited to): negative controls, positive reference substance；It can also include quantitative fluorescent PCR
Reaction plate, PCR reaction plate sealed membrane etc..
Further, the chip includes: solid phase carrier, and the oligonucleotides being orderly fixed on the solid phase carrier is visited
Needle, the oligonucleotide probe specifically correspond to some or all of above-mentioned LncRNA marker sequence.Wherein, solid phase carries
Body includes sheet glass, silicon wafer, polypropylene screen, nitrocellulose filter, nylon membrane or polystyrene film.
The present invention provides a kind of application of the said goods in the tool of preparation diagnosis diabetes.Reagent in the present invention
Box or chip can be used for detecting multiple genes including NONHSAT024449 (for example, multiple bases relevant to diabetes
Cause) expression, by multiple markers of diabetes simultaneously detect, be greatly improved the accuracy rate of diabetes diagnosis.
Detection in the present invention includes but is not limited to sequencing technologies, nucleic acid hybridization technique, nucleic acid amplification technologies or immunoassays, core
Sour amplification technique includes but is not limited to polymerase chain reaction, reverse transcriptase polymerase chain reaction, the amplification of transcriptive intermediate, connection
Enzyme chain reaction, strand displacement amplification or the amplification based on nucleic acid sequence.
Compared with prior art, the beneficial effects of the present invention are:
Present invention firstly discovers that LncRNA molecular marked compound NONHSAT024449 have in diabetic's blood it is brighter
Aobvious differential expression, compared with normal population control group, diabetic population experimental group patient has the LncRNA molecule of low expression
Marker NONHSAT024449, is based on this discovery, and LncRNA molecular marked compound NONHSAT024449 can be used as diabetes point
Sub- marker or target spot are applied to diabetes clinical diagnosis or targeted therapy, by detecting LncRNA molecule provided by the present invention
The specific primer of marker NONHSAT024449 is to early screening, the prevention/assessment diabetes hair that diabetes may be implemented
Raw risk is applied to LncRNA molecular marked compound NONHSAT024449 to prepare diabetes diagnosis product or preparation diabetes is pre-
In anti-/ assessment product, be conducive to further elucidate diabetes occurrence and development mechanism, signal path etc., great application prospect and reason
Value.
Detailed description of the invention
Fig. 1 is that the QPCR of NONHSAT024449 marker expands electrophoretogram；M is DNA molecular amount standard, and number 1-5 is represented
Diabetic population individual, number 6 represent negative control in experimental group；
Fig. 2 is the relative expression quantity situation map of diabetic population and normal population blood NONHSAT024449 in embodiment 2.
Specific embodiment
With reference to the accompanying drawing, the present invention is further illustrated, these embodiments are merely to illustrate the present invention rather than limitation
The scope of the invention；Test method without specific conditions in embodiment, according to normal conditions, examination used in embodiment
The amount of agent box reagent is merely exemplary, and those skilled in the art can be adjusted accordingly according to the actual situation；The reagent and
Biomaterial commercially obtains unless otherwise specified.In the present invention, " probe " refer to can be with the spy of another molecule
The molecule that sequencing column or subsequence or other parts combine.Unless otherwise indicated, term " probe " is often referred to that complementary base can be passed through
The polynucleotide probes that basigamy pair is combined with another polynucleotides (often referred to as " target polynucleotide ").According to the tight of hybridization conditions
Careful property, probe energy and with the probe lack sufficient sequence complementarity target polynucleotide in conjunction with.Probe can be made direct or indirect
Label, range includes primer.Crossing system includes, but are not limited to: solution phase, solid phase, mixed phase or in situ hybridization measurement
Method.
The probe has the base sequence with the specific base sequence complementary of target gene.Here, so-called " complementation ", as long as
It is hybridization, can not be complete complementary.These polynucleotides usually relative to the specific base sequence have 80% with
Upper, preferably 90% or more, more preferable 95% or more, particularly preferred 100% homology.These probes can be DNA, can also be with
It is RNA, furthermore it is possible to pass through PNA (peptide nucleic acid), LNA (Cross-linked nucleic acid), ENA in part of it or whole nucleotides
The polynucleotides that the artificial replacement nucleic acid such as (glycerol nucleic acid), TNA (threose nucleic acid) obtains.
It would be recognized by those skilled in the art that practicability of the invention is not limited to any of marker gene of the invention
The gene expression of specific variants is quantified.As unrestricted example, marker gene can have SEQ ID NO.1 to refer to
Fixed sequence.In some embodiments, have with the same or similar cDNA sequence of listed sequence at least 85%, such as
Above-mentioned listed sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least 99% are identical
Or similar cDNA sequence.
In specific embodiments of the present invention, the reduction of NONHSAT024449 gene level is usually compared with the control
Reduction.Technical staff can select maximally related control.This is also generally dependent on the property of institute's study of disease, obtainable sample
Product etc..Suitable control includes but is not limited to, the average water of the similar sample of the subject from non-diabetic, control group
Flat or one group of clinical data in relation to gene NONHSAT024449 product average level in sampling tissue.From it is above-mentioned can be clearly
Find out, control can come from identical subject or from one or more different subjects or from clinical data.It is optional
Ground, to impinge upon such as gender, the age on match.Test method without specific conditions in embodiment, according to routine
Condition；The reagent and biomaterial commercially obtain unless otherwise specified.
Embodiment 1
1. sampling: acquire the anticoagulated blood 5mL of 107 Xue Tang≤7.1mmol/L diabetic populations (experimental group) respectively, 107
It does not suffer from and send in normal population (control group) the anticoagulated blood 5mL, 1h of diabetes and other diseases to laboratory, all samples
Patient's informed consent before obtaining, and pass through the agreement of the committee, organizational ethics.3 will be added in the sample of above-mentioned acquirement
The erythrocyte cracked liquid of times whole blood is stored at room temperature 10min after being mixed by inversion, 3000rpm is centrifuged 5min, abandons supernatant, is precipitating
In 2mL erythrocyte cracked liquid is added again, be stored at room temperature 10min after mixing, abandon supernatant after 3000rpm centrifugation 5min.It is precipitating
Middle addition 2mL Trizol dissolution, is blown and beaten not sticky to solution using pipette tips, deposits in -80 DEG C of refrigerators rapidly.
2. the extraction of leucocyte total serum IgE sample:
(1) chloroform of 1mL is added, be vortexed concussion at least 30s, and 13000rpm is centrifuged 5min；
(2) it takes supernatant in new EP pipe, 0.8mL isopropanol is added, mitigate and mix, be incubated at room temperature at least 10min,
13000rpm is centrifuged 5min；
(3) supernatant is abandoned, 75% dehydrated alcohol of 2mL is added, abundant vortex mixes, and 13000rpm is centrifuged 5min；
(4) supernatant is abandoned, dries precipitating with room temperature；
(5) 25-30 μ L is added in precipitating without enzyme water (RNase free water), in 4 DEG C dissolution 1-2 hours, in 56 DEG C of gold
Belong in bath and be incubated for 10min, RNA enzyme is inactivated.
3. reverse transcription polymerase chain reaction (RT-PCR)
The RNA extracted in step 2 is added 1000ng RNA by 10 μ L systems and determines sample rna additional amount, i.e. and sample additional amount X=
(1000/RNA concentration) × (reaction system/10)；(1) reaction system: 5 × PrimeScript RT mixed liquor, 2 μ L, without enzyme water
(RNase free water) 7 μ L, 1 μ L, RT-PCR total volume of RNA sample, 10 μ L.
(2) RT-PCR reaction condition: 37 DEG C of 15min, 85 DEG C of 5s, 4 DEG C of long-term heat preservations.Gained reverse transcription cDNA product is vertical
It reacts for qPCR, or is saved under -80 DEG C of environment, use finishes in half a year, and multigelation should be avoided in cDNA.
4. quantitative PCR (Q-PCR)
(1) Q-PCR reaction system: 10 μ L of TB Green Premix Ex Taq, upstream primer (Foward primer, 10 μM)
0.4 μ L, downstream primer (Reverse primer, 10 μM) 0.4 μ L, dyestuff (ROX Reference Dye/ Dye II) 0.4 μ
L, 2 μ L of reverse transcription cDNA template, without enzyme water (RNase free water) be added to 20 μ L.Its upstream primer nucleotide sequences is such as
In sequence table described in SEQ ID NO.2, downstream primer nucleotide sequence is as described in SEQ ID NO.3 in sequence table.
(2) Q-PCR reaction condition is as follows: 95 DEG C of 5min of initial denaturation are recycled 1 time, 95 DEG C of 15s, 60 DEG C of 40s, are recycled 45 times,
Fluorescence is detected at 60 DEG C.
Using diabetic population experimental group as positive control, blank control is that negative control carries out PCR detection, PCR amplification knot
For fruit as shown in Figure 1, in figure, M is DNA molecular amount standard, and number 1 ~ 5 represents diabetic population individual in experimental group, and number 6 represents
Blank control；There is an apparent characteristic purpose band at 271bp in experimental group, and blank control there are no the DNA of 271bp
Band.
Embodiment 2: expression of the marker LncRNA in diabetic tissue
It is fixed in Light Cycler fluorescence according to the PCR amplification of embodiment 1 as a result, using SYBR Green as fluorescent marker
It measures and carries out PCR reaction in PCR instrument, determine that purpose band, Δ Δ CT method carry out relative quantification by melt curve analysis analysis and electrophoresis.
It is repeated 3 times test, result data is indicated in a manner of mean+SD, using SPSS18.0 statistical software
Come for statistical analysis, the paired comparisons of normal population control group and diabetic population experimental group are using t inspection, it is believed that when P <
There is statistical significance when 0.05.As a result as shown in Fig. 2, NONHSAT024449 gene is in diabetic population experimental group blood
Expression quantity is significantly lower than normal population control group, and low expression amount is presented, and illustrates that the two has apparent negatively correlated relationship, has
Statistical significance (P < 0.05).
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Sequence table
<110>Xuzhou Municipal Disease Control and Prevention Center (Institute of Health Education, Xuzhou City)
<120>a kind of LncRNA marker and its application in diabetes
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2435
<212> DNA
<213>source of people (Homo sapiens)
<400> 1
gaggcagagt ctcactctgt cgccaggctg gagtgcagtg gtgtgatctt ggctcactgc 60
aacctctgcc tcctgggttc aagcgattct tgtgcctcag cctctcgagt agctgggatt 120
acaggcatgc gccaccacac ccagctaatt tttgtgtttt tagtagagac ggggtttcac 180
cattttggcc aggatggtct caatcccctg acctcgtgat ccacctgcct cggcctccca 240
aagtgttggg attacaggca tgagccactg tgcttggcct gttattttat tttcttataa 300
ctacaacttt tcttcttgaa ttttcaggtc agaggcaaga aaaactcttt acaggttttt 360
agtggggggc ttatggagta tttcaggagt tctttgcaaa ttaaatcatc ttttcacttg 420
tattgttttt caaaactttg ttgatttcta aaatgtgcca actgtgagta aactatggta 480
tttgcaagtg gtttttacat aatatttgag atgaggaagt gagattgtgc atgacatact 540
tctcctttgt attctctcag tgccttacag caggttactc cattctgcta tgacaacttg 600
tttcaaatgt taatttacat aggatttttt ataagccatt aaggcatatg tatagtatat 660
cagtaaagat ggatggtgca tatataaata gtcttctgta atagtgattg gatttacttc 720
tcaattatga gagacaaaaa ttatcccctc acctgtctct attctttcaa caggttgatc 780
ccttttcatg atttttcatt aggtggttca ggaagtttcc atattacagc gcttcagact 840
gtatatgtta gtttaaaaat cacttttctc tctctcaact tctttctttt ttttttgaag 900
acttaattta aaaaatttgg gttgttagat ccgtatcata gatttggcct agcctcttct 960
gttaacctag tccacagatg agcgaatctg gttagttgaa ggacattgtg atttgactct 1020
ggtcacgcga ggaagtagaa gggcaaagac aggaccggca gtttacattt ccagtggtta 1080
aacctcacgg tactttggga ctgcttgtta acttttgtgg ttgtctgagg ccaatctaac 1140
gtgaccattt ctgacacctc aacagagaga ggaaagcaac ttgagcaatg agagtaaata 1200
acttgggctc tcagagattt gaagatagag atctcattgt gagggggact attttgcagg 1260
tcctcatttc tccaagaaag agatggtgtt acaggaaccc actgaaagcc atatcccatt 1320
aaatgaggaa ctaattttgg ctgggccttc ttgtaatgtc ctcgcaggtg tgttgtgaag 1380
attaatgcag ggtagtatgt ttgtagattg acacctagtc taaacttgag gtaattggtg 1440
ctctgtgaat actcagtcgt gttcttttat agccttaatc atgatttgaa ctagtccctt 1500
gctttttaaa tgactgaatg aagtccttcg tggtaaggga gtacgttgat aacttagttt 1560
actatatggg tttgtggtcg catcccagtc atcagctgct atcattttcc ttcttcatcc 1620
cttatactga gatttgggtt acagcttttt attcttcgaa ggatcacaaa gcagtgtaca 1680
gacacctgcc ttctttaagg atgaaaggaa gataaagtgg tctttttttg tttacttatt 1740
tgtttcacct cttgtttgag taacttctaa ggtgctattc tctctctctt tttgctacct 1800
catgagctct tgtcacagcc atggaaacca gcctcgttta gaaagggaac ttagttcaga 1860
aggggttaaa agccttccag aatttttctt tagctgctga agtttttaca tgtggttaca 1920
tgactttaag ttttatgcat tacgctctta attctattac aaaatgtgga ctcaccaatt 1980
gctttgtgtt ttccatgtga cctgttactt caggctactt ggggaacatc ttagtcctct 2040
gtagctcctg aacccagcac tggtgcttca agagagaagg tagcacgtct ttgttcaaaa 2100
caaaacaaaa cgacacttct ggaggccaca tcctgaatat gaatgttcta ctaagtcact 2160
cagttatggt tctaaaggga aactgtaaga agacccacaa ggagtggacc aagactatta 2220
tttaattgca caacttgaaa ctttgctgcc agaagaggca gctccattcc tttgactcca 2280
gtgttgggct gttaactgct gcacctcatt gccttttttt gtttttgttt ttgttttgta 2340
ggagggtagg cactgttggg ccatatgcac aaatattgta actcttggta tctttactgc 2400
atcatagtca ataaacttct ttgtaccctt tggct 2435
<210> 2
<211> 18
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
tcaggtcaga ggcaagaa 18
<210> 3
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
gtcatagcag aatggagtaa c 21
Sequence table
<110>Xuzhou Municipal Disease Control and Prevention Center (Institute of Health Education, Xuzhou City)
<120>a kind of LncRNA marker and its application in diabetes
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2435
<212> DNA
<213>source of people (Homo sapiens)
<400> 1
gaggcagagt ctcactctgt cgccaggctg gagtgcagtg gtgtgatctt ggctcactgc 60
aacctctgcc tcctgggttc aagcgattct tgtgcctcag cctctcgagt agctgggatt 120
acaggcatgc gccaccacac ccagctaatt tttgtgtttt tagtagagac ggggtttcac 180
cattttggcc aggatggtct caatcccctg acctcgtgat ccacctgcct cggcctccca 240
aagtgttggg attacaggca tgagccactg tgcttggcct gttattttat tttcttataa 300
ctacaacttt tcttcttgaa ttttcaggtc agaggcaaga aaaactcttt acaggttttt 360
agtggggggc ttatggagta tttcaggagt tctttgcaaa ttaaatcatc ttttcacttg 420
tattgttttt caaaactttg ttgatttcta aaatgtgcca actgtgagta aactatggta 480
tttgcaagtg gtttttacat aatatttgag atgaggaagt gagattgtgc atgacatact 540
tctcctttgt attctctcag tgccttacag caggttactc cattctgcta tgacaacttg 600
tttcaaatgt taatttacat aggatttttt ataagccatt aaggcatatg tatagtatat 660
cagtaaagat ggatggtgca tatataaata gtcttctgta atagtgattg gatttacttc 720
tcaattatga gagacaaaaa ttatcccctc acctgtctct attctttcaa caggttgatc 780
ccttttcatg atttttcatt aggtggttca ggaagtttcc atattacagc gcttcagact 840
gtatatgtta gtttaaaaat cacttttctc tctctcaact tctttctttt ttttttgaag 900
acttaattta aaaaatttgg gttgttagat ccgtatcata gatttggcct agcctcttct 960
gttaacctag tccacagatg agcgaatctg gttagttgaa ggacattgtg atttgactct 1020
ggtcacgcga ggaagtagaa gggcaaagac aggaccggca gtttacattt ccagtggtta 1080
aacctcacgg tactttggga ctgcttgtta acttttgtgg ttgtctgagg ccaatctaac 1140
gtgaccattt ctgacacctc aacagagaga ggaaagcaac ttgagcaatg agagtaaata 1200
acttgggctc tcagagattt gaagatagag atctcattgt gagggggact attttgcagg 1260
tcctcatttc tccaagaaag agatggtgtt acaggaaccc actgaaagcc atatcccatt 1320
aaatgaggaa ctaattttgg ctgggccttc ttgtaatgtc ctcgcaggtg tgttgtgaag 1380
attaatgcag ggtagtatgt ttgtagattg acacctagtc taaacttgag gtaattggtg 1440
ctctgtgaat actcagtcgt gttcttttat agccttaatc atgatttgaa ctagtccctt 1500
gctttttaaa tgactgaatg aagtccttcg tggtaaggga gtacgttgat aacttagttt 1560
actatatggg tttgtggtcg catcccagtc atcagctgct atcattttcc ttcttcatcc 1620
cttatactga gatttgggtt acagcttttt attcttcgaa ggatcacaaa gcagtgtaca 1680
gacacctgcc ttctttaagg atgaaaggaa gataaagtgg tctttttttg tttacttatt 1740
tgtttcacct cttgtttgag taacttctaa ggtgctattc tctctctctt tttgctacct 1800
catgagctct tgtcacagcc atggaaacca gcctcgttta gaaagggaac ttagttcaga 1860
aggggttaaa agccttccag aatttttctt tagctgctga agtttttaca tgtggttaca 1920
tgactttaag ttttatgcat tacgctctta attctattac aaaatgtgga ctcaccaatt 1980
gctttgtgtt ttccatgtga cctgttactt caggctactt ggggaacatc ttagtcctct 2040
gtagctcctg aacccagcac tggtgcttca agagagaagg tagcacgtct ttgttcaaaa 2100
caaaacaaaa cgacacttct ggaggccaca tcctgaatat gaatgttcta ctaagtcact 2160
cagttatggt tctaaaggga aactgtaaga agacccacaa ggagtggacc aagactatta 2220
tttaattgca caacttgaaa ctttgctgcc agaagaggca gctccattcc tttgactcca 2280
gtgttgggct gttaactgct gcacctcatt gccttttttt gtttttgttt ttgttttgta 2340
ggagggtagg cactgttggg ccatatgcac aaatattgta actcttggta tctttactgc 2400
atcatagtca ataaacttct ttgtaccctt tggct 2435
<210> 2
<211> 18
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
tcaggtcaga ggcaagaa 18
<210> 3
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
gtcatagcag aatggagtaa c 21
Claims (9)
1. a kind of LncRNA marker, which is characterized in that the LncRNA marker is NONHSAT024449, described
The nucleotide sequence of LncRNA marker is as shown in SEQ ID NO.1.
2. marker according to claim 1, which is characterized in that the specific primer pair of the LncRNA marker,
Upstream primer nucleotide sequence is as shown in SEQ ID NO.2, and primer nucleotide sequences are as shown in SEQ ID NO.3 downstream.
3. the application of LncRNA marker as described in claim 1, which is characterized in that the LncRNA marker is assessed in preparation
Or the application in the product of diagnosis diabetes.
4. application according to claim 3, which is characterized in that the product passes through RT-PCR, real-time quantitative PCR, original position
Hybridization, chip or high-flux sequence method detection claim 1 described in LncRNA marker expression to assess or
Diagnose diabetes.
5. the product of NONHSAT024449 content in a kind of measurement sample, which is characterized in that the product is in preparation assessment or examines
Application in the tool of disconnected diabetes.
6. product according to claim 5, which is characterized in that the product includes kit, probe, chip or preparation.
7. product according to claim 6, which is characterized in that the chip includes: solid phase carrier, and is orderly fixed on
Oligonucleotide probe on the solid phase carrier, the oligonucleotide probe specifically correspond to described in claim 1
Some or all of LncRNA marker sequence.
8. product according to claim 6, which is characterized in that containing as claimed in claim 2 in institute's kit
The specific primer pair of LncRNA marker.
9. product according to claim 6, which is characterized in that the kit further includes dNTP, random primer, reduction
Agent, RNase inhibitor, reverse transcriptase, MgCl2With one of PCR buffer etc. or a variety of combinations.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112646879A (en) * | 2021-01-25 | 2021-04-13 | 中国药科大学 | Application of non-coding RNA biomarker lnc-Wdr5 |
| CN113215242A (en) * | 2021-04-23 | 2021-08-06 | 皖南医学院第一附属医院（皖南医学院弋矶山医院） | Molecular marker of type 2diabetes and application thereof |
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|---|---|---|---|---|
| WO2015092020A2 (en) * | 2013-12-20 | 2015-06-25 | Université de Lausanne | Diagnostic, prognostic and therapeutic uses of long noncoding rnas for heart disease and regenerative medicine |
| CN105543400A (en) * | 2016-02-29 | 2016-05-04 | 北京泱深生物信息技术有限公司 | Molecular marker of diabetes mellitus type I |
| CN106434872A (en) * | 2016-08-11 | 2017-02-22 | 河南大学 | MiRNA molecule marker hsa-miR-152-3p for diagnosing type 2 diabetes, and application thereof |
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2019
- 2019-05-20 CN CN201910419178.2A patent/CN110396542A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015092020A2 (en) * | 2013-12-20 | 2015-06-25 | Université de Lausanne | Diagnostic, prognostic and therapeutic uses of long noncoding rnas for heart disease and regenerative medicine |
| CN105543400A (en) * | 2016-02-29 | 2016-05-04 | 北京泱深生物信息技术有限公司 | Molecular marker of diabetes mellitus type I |
| CN106434872A (en) * | 2016-08-11 | 2017-02-22 | 河南大学 | MiRNA molecule marker hsa-miR-152-3p for diagnosing type 2 diabetes, and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112646879A (en) * | 2021-01-25 | 2021-04-13 | 中国药科大学 | Application of non-coding RNA biomarker lnc-Wdr5 |
| CN112646879B (en) * | 2021-01-25 | 2022-11-04 | 中国药科大学 | Application of non-coding RNA biomarker lnc-Wdr5 |
| CN113215242A (en) * | 2021-04-23 | 2021-08-06 | 皖南医学院第一附属医院（皖南医学院弋矶山医院） | Molecular marker of type 2diabetes and application thereof |
| CN113215242B (en) * | 2021-04-23 | 2022-05-31 | 皖南医学院第一附属医院（皖南医学院弋矶山医院） | Molecular marker for type 2diabetes and application thereof |
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