CN106676109A - ENST00000418539.1, preparation or diagnostic agent or medicine or kit, and application of ENST00000418539.1 - Google Patents
ENST00000418539.1, preparation or diagnostic agent or medicine or kit, and application of ENST00000418539.1 Download PDFInfo
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Abstract
The invention relates to the technical field of gene engineering, and relates to a long-chain non-coding RNA ENST00000418539.1, a preparation or a diagnostic agent or a medicine or a kit, a detection method, and application of the ENST00000418539.1. The sequence of the long-chain non-coding RNA ENST00000418539.1 is shown as SEQ ID NO:1 in the description. The long-chain non-coding expression profile chip technology is utilized, the long-chain non-coding RNA ENST00000418539.1, which is remarkably highly expressed in the blood of a stable angina pectoris patient, is screened through differential analysis, and the researches on the pathogenesis of the stable angina pectoris are further enriched.
Description
Technical field
The present invention relates to gene engineering technology field, is a kind of long-chain non-coding RNA ENST00000418539.1 and system
Agent or diagnostic agent or medicine or test kit and its method and its application is detected, be i.e. ENST00000418539.1 and preparation or examined
Disconnected agent or medicine or test kit and application.
Background technology
Stable angina pectoris (Stable angina, SA) are a kind of cardiovascular disease of threat human health, although its is pre-
It is good afterwards, but still have some patientss some months after making a definite diagnosis in several years, the painstaking effort such as acute myocardial infarction and apoplexy to occur and runs affairs
Part, even it is dead, seriously threaten life and the quality of life of people.Although the disease has controlling including numerous including Drug therapy
Treatment means, occupy every year huge medical resource, although the progress of medical procedure and the increasing of input, the sickness rate and disease of SA
Dead rate still remains high.The early diagnosiss of SA have important value for pre- angina pectoris develop into clinical cardiovascular events, can
To improve the survival rate of patient with angina pectoris and long-term prognosis can be improved.Therefore, searching can early diagnosiss SA patient and have compared with
The new biomarker of hypersensitivity and specificity seems particularly necessary.
Long-chain non-coding RNA (long non-coding RNAs, lncRNA) is a class transcript length more than 200nt
RNA molecule, typically not encoding proteins, by rna plymerase ii transcription and Jing variable sheers are formed, transcriptional level is compiled less than protein
Code gene, had once been considered as once " noise " of subgenomic transcription.It has proven convenient that long-chain non-coding RNA is in epigenetics, transcription
Various aspect controlling gene expression such as regulation and control, post-transcriptional control, play a significant role in various biological process, including fat
Generation of metabolism, atherogenesis, fetal development and tumor etc..
The content of the invention
The invention provides the preparation or diagnostic agent or medicine of a kind of long-chain non-coding RNA ENST00000418539.1 or
Test kit, overcomes the deficiency of above-mentioned prior art, and the early diagnosiss of its energy effectively solving stable angina pectoris are for the prevention heart
Angor develops into clinical cardiovascular events and has important value, can improve the survival rate of patient with angina pectoris and can improve long-term
Prognosis, find can early diagnosiss patients with stable angina pectoris and have compared with hypersensitivity and specificity new biomarker show
Particularly necessary problem is obtained, present invention discover that its expression or dysfunction and the close phase of the occurrence and development of human cardiovascular disease
Close, long-chain non-coding RNA is expected to become cardiovascular disease diagnosis, the mark of prediction prognosis, while also controlling for cardiovascular diseasess
Treat and provide new target spot.
One of technical scheme is realized by following measures：One kind is in patients with stable angina pectoris blood
The long-chain non-coding RNA ENST00000418539.1 of middle high expression, long-chain non-coding RNA ENST00000418539.1's
Sequence such as SEQ ID NO:Shown in 1.
The further optimization and/or improvements to one of foregoing invention technical scheme are presented herein below：
Above-mentioned long-chain non-coding RNA ENST00000418539.1 is ENST00000418539.1:1 total length or piece
Section.
The two of technical scheme are realized by following measures：The non-volume of long-chain in a kind of vitro detection blood
The preparation or diagnostic agent or medicine or test kit of code RNA ENST00000418539.1, said preparation or diagnostic agent or medicine or examination
Agent box include specific primer to, standard DNA template, PCR reactant liquors, wherein, specific primer to including forward primer and under
Trip primer, the nucleotides sequence of forward primer is classified as SEQ ID No:2, the nucleotides sequence of downstream primer is classified as SEQ ID No:3.
To foregoing invention technical scheme two further optimization and/or improvements are presented herein below：
The above-mentioned test kit is fluorescent quantificationally PCR detecting kit.
Above-mentioned specific primer is to for SYBR Green, Taqman probes, molecular beacon, double cross probe, compound spy
The detection of pin.
Above-mentioned PCR reactant liquors are fluorescence quantitative PCR reaction solution.
Above-mentioned fluorescence quantitative PCR reaction solution includes dNTP, Mg2+, Taq enzyme.
It is above-mentioned also to include fluorescent dye.
The three of technical scheme are realized by following measures：One kind detection is according to one of technical scheme institute
The method of the long-chain non-coding RNA ENST00000418539.1 for stating, is carried out in the steps below：The first step, extracts blood total
RNA；Second step, prepares cDNA；3rd step, quantitative amplification long-chain non-coding RNA ENST00000418539.1.
The four of technical scheme are realized by following measures：It is a kind of according to one of technical scheme
Long-chain non-coding RNA ENST00000418539.1 is steady in the test kit prepared for detecting stable angina pectoris or as detection
The application of the novel targets of sizing angina drug.
The present invention utilizes long-chain non-coding chip of expression spectrum technology, by variation analyses, screens one in the stable type heart
The long-chain non-coding RNA ENST00000418539.1 of significantly high expression in angor blood samples of patients, further enriches the stable type heart
The research of angor pathogenesis, meanwhile, vitro detection stable angina pectoris related gene of the present invention is long-chain non-coding
The reagent of RNA ENST00000418539.1 and vitro detection stable angina pectoris related gene of the present invention are long-chain
The test kit of non-coding RNA ENST00000418539.1 can be used in long-chain non-coding RNA of the present invention
The detection of the expression of ENST00000418539.1, by long-chain non-coding RNA of the present invention
The testing result of the expression of ENST00000418539.1 knows the screening results of stable angina pectoris, is that the stable type heart is twisted
The diagnosis and treatment of pain provides new direction.
Description of the drawings
Accompanying drawing 1 detects ENST00000418539.1 in normal healthy controls for long-chain non-coding RNA chip of expression spectrum in the present invention
Organize and the detected signal value in stable angina pectoris group.
Accompanying drawing 2 is special a pair of the sequential design in the present invention for long-chain non-coding RNA ENST00000418539.1
Row agarose gel electrophoresis test the effect of primer after specific primer PCR amplifications.
Accompanying drawing 3 is qRT-PCR detection RNA ENST00000418539.1 in the present invention in healthy control group and the stable type heart
Relative expression quantity in angor group.
Specific embodiment
The present invention is not limited by following embodiments, can technology according to the present invention scheme and practical situation determining specifically
Embodiment.
In the present invention, for the ease of description, the description of the relative position relation of each part is according to Figure of description 1
Butut mode being described, such as：The position relationship of upper and lower, left and right etc. is come according to the Butut direction of Figure of description
It is determined that.
The present invention utilizes long-chain non-coding chip of expression spectrum technology, by variation analyses, screens one in the stable type heart
The long-chain non-coding RNA ENST00000418539.1 of significantly high expression in angor blood samples of patients, its transcript regions is positioned at No. 2 dyes
Colour solid, original position is 47,558,199 to 47,571,656, total length 13,458bp.It is long compared with the blood content of Healthy People
Chain non-coding RNA ENST00000418539.1 significantly high expression in patients with stable angina pectoris blood, and in the glimmering of sample
Long-chain non-coding RNA ENST00000418539.1 is further characterized by light quantitative experiment in patients with stable angina pectoris blood
It is significantly higher than Healthy People.This new long-chain non-coding RNA ENST00000418539.1 will further enrich the stable type heart
The research of angor pathogenesis, also the early diagnosiss and Prognosis scoveillance for stable angina pectoris provide new blood molecules mark
Thing and therapy target.
The patients with stable angina pectoris and the blood preparation of Healthy People that inventor provides is each 5, totally 10.It is public by ABI
Step needed for the Trizol reagents (article No. 15596-026) of department is extracted after total serum IgE, using Beijing Bo Ao biological engineering company limited
Chip product (Agilent human lncRNA+mRNA Array V4.0) detected, filter out one in the stable type heart
The long-chain non-coding RNA ENST00000418539.1 of significantly high expression in angor blood samples of patients, its nucleotide sequence such as SEQ ID
Shown in No.1.Later stage through carrying out quantitative fluorescent PCR checking to sample, it is found that patients with stable angina pectoris is notable in 10 samples
Higher than Healthy People.Long-chain non-coding RNA ENST00000418539.1, as the novel targets of stable angina pectoris, is the stable type heart
The clinical treatment and drug development of angor is provided fundamental basis.
Embodiment 1, a kind of long-chain non-coding RNA of expression high in patients with stable angina pectoris blood
The sequence such as SEQ ID NO of ENST00000418539.1, long-chain non-coding RNA ENST00000418539.1:Shown in 1.Should
Long-chain non-coding RNA ENST00000418539.1 can enrich the research of stable angina pectoris pathogenesis, and it can be carried out
Early diagnosiss.
Embodiment 2, used as the optimization of embodiment 1, long-chain non-coding RNA ENST00000418539.1 is
ENST00000418539.1:1 total length or fragment.
Embodiment 3, the long-chain non-coding RNA in a kind of vitro detection blood according to embodiment 1 and embodiment 2
The preparation or diagnostic agent of ENST00000418539.1 or medicine or test kit, said preparation or diagnostic agent or medicine or test kit bag
Include specific primer to, standard DNA template, PCR reactant liquors, wherein, specific primer to including forward primer and downstream primer,
The nucleotides sequence of forward primer is classified as SEQ ID No:2, the nucleotides sequence of downstream primer is classified as SEQ ID No:3.The long-chain is non-
The preparation or diagnostic agent of coding RNA ENST00000418539.1 or medicine or test kit can be twisted more easily to the stable type heart
The blood serum designated object of pain is diagnosed and predicted.
Embodiment 4, used as the optimization of embodiment 3, the test kit is fluorescent quantificationally PCR detecting kit.
Embodiment 5, used as the optimization of embodiment 3 or embodiment 4, specific primer is to for SYBR Green, Taqman
Probe, molecular beacon, double cross probe, the detection of combined probe.
Embodiment 6, used as the optimization of embodiment 3, embodiment 4 and embodiment 5, PCR reactant liquors are quantitative fluorescent PCR reaction
Liquid.
Embodiment 7, used as the optimization of embodiment 6, fluorescence quantitative PCR reaction solution includes dNTP, Mg2+, Taq enzyme.
Embodiment 8, used as the optimization of embodiment 3, embodiment 4, embodiment 5, embodiment 6 and embodiment 7, the test kit is also
Including fluorescent dye.
The using method of the dye class PCR kit for fluorescence quantitative of the vitro detection stable angina pectoris, quantitative fluorescent PCR
System：
Forward primer, downstream primer each 0.25ul (10uM)；DNA profiling cDNA 0.5ul；Power SYBR Green
PCR Master (2 ×) 5ul, plus Nuclease-Free Water4ul, total capacity 10ul.Quantitative fluorescent PCR program：The first step
95 DEG C 10 minutes；2nd step, 95 DEG C of 15S, 60 DEG C 1 minute；3rd step, 95 DEG C of 15S, 60 DEG C 1 minute, 95 DEG C of 15S, totally 40 are followed
Ring.
Embodiment 9, a kind of long-chain non-coding RNA of detection according to embodiment 1 and embodiment 2
The method of ENST00000418539.1, is carried out in the steps below：The first step, extracts blood total serum IgE；Second step, prepares cDNA；
3rd step, quantitative amplification long-chain non-coding RNA ENST00000418539.1.
Methods described specifically includes following steps：
First, blood total serum IgE is extracted：According to reagent and step needed for the Trizol reagents (article No. 15596-026) of ABI companies
Total serum IgE is extracted, then it is fixed with 7300real time PCR system nucleic acid quantifications instrument quantitative (Applied Biosystems AB)
The extracted purity of amount and concentration.
2nd, sample cDNA is prepared：Using Beijing Tiangeng biochemical technology company FastQuant cDNA the first chain synthetic agents
Box test kit (article No. KR106) synthesizes cDNA to the total serum IgE reverse transcription extracted.
1st, template ribonucleic acid is thawed on ice；5×gDNA Buffer、FQ-RT Primer Mix、10×Fast RT
Buffer、RNase-Free ddH2O is immediately placed on ice in (15-25 DEG C) defrosting of room temperature after defrosting.Using it is front will be every kind of molten
Liquid vortex oscillation is mixed, and brief centrifugation remains in the liquid of tube wall to collect.
2nd, secondary structure is opened, reaction system and condition are as shown in table 1；
The removal system of the genomic DNA of said components is prepared into mixed liquor, is thoroughly mixed.Brief centrifugation, is placed in 42
DEG C, it is incubated 3min.It is subsequently placed in and places on ice.
3rd, reverse transcription reaction, reaction system and condition it is as shown in table 2；
By the Mix in reverse transcription reaction, in being added to the reactant liquor of gDNA removal steps, fully mix.42 DEG C, incubation
15min.95 DEG C, it is put in the cDNA on ice, obtaining after incubation 3min and can be used for subsequent experimental, or cryopreservation.
3rd, long-chain non-coding RNA ENST00000418539.1 is expanded：Using the Power SYBR Green of ABI companies
PCR Master Mix fluorescence quantitative kits (article No. 4367659), by template of the cDNA of reverse transcription quantitative fluorescent PCR is carried out
Amplification.
Quantitative fluorescent PCR system is as shown in table 3；
Quantitative fluorescent PCR program：The first step 95 DEG C 10 minutes；2nd step, 95 DEG C of 15S, 60 DEG C 1 minute；3rd step, 95 DEG C
15S, 60 DEG C 1 minute, 95 DEG C of 15S, totally 40 circulation.
Embodiment 10, a kind of long-chain non-coding RNA according to embodiment 1 and embodiment 2
ENST00000418539.1 is in the test kit prepared for detecting stable angina pectoris or as detection stable angina pectoris medicine
Novel targets application.
Method for detecting stable angina pectoris, the method comprising the steps of：
The first step, extracts blood total serum IgE；
Second step, prepares sample cDNA；
3rd step, expands long-chain non-coding RNA ENST00000418539.1, and is sentenced according to relative quantification result
It is disconnected.
Embodiment 11：Patients with stable angina pectoris and the long-chain non-coding RNA chip expression analysis of healthy human blood.
First, material and method
1. material
It is each 5 with healthy human blood's sample that blood sample comes from patients with stable angina pectoris.
2. method
(1) extraction of patients with stable angina pectoris and healthy human blood's sample total serum IgE：According to the limited public affairs of rich biotechnology difficult to understand
" for the preparation process of the blood sample of gene microarray analysis " that department provides extracts patients with stable angina pectoris with healthy human blood
Liquid sample total serum IgE.Specific experiment material includes：Lymphocyte separation medium (Tianjin TBD biotech developments center)；Sterile physiological
Saline (common 0.9% normal saline sterilizing)；Trizol reagents (Invitrogen companies).
(2)LncRNA+mRNA chip of expression spectrum is detected with the total serum IgE of detected sample as starting, expanded in vitro
Increase and fluorescent labeling, labeling process is adoptedBiochip common tags test kit.Mainly comprise the steps：
1) reverse transcription synthesizes the first chain cDNA：It is initial with Total RNA, the T7Oligo containing T7 promoter sequences (dT)
Pr imer and T7- random primers, both can be with the mRNA of junction belt poly (A), it is also possible to combine in addition to rRNA other without
The RNA of poly (A), using the first chain Enzyme Mix the first chain cDNA is synthesized.
2) the second chain DNA is synthesized：The RNA chains in DNA-RNA heterozygotes are converted into into second with the second chain Enzyme Mix
Chain cDNA, synthetic dsdna.
3) in vitro transcription synthesis cRNA：With the second chain cDNA as template, using T7Enzyme Mix cRNA is synthesized.
4) cRNA purification：Using RNA purification column purification cRNA, except reagents such as the salt in dereaction, enzymes, and cRNA is carried out
Quantitatively, Quality Control.
5) reverse transcription：With cRNA as template, Random Primer are primer, and the enzymes of CbcScript II carry out reverse transcription.It is pure
The cDNA that change recovery reverse transcription is obtained is simultaneously quantitative.
6) labelling：As template, Random Primer are primer to cDNA products with reverse transcription, use Klenow Fragment
Enzymatic synthesiss cDNA complementary strands simultaneously mix the dNTP (Cy3-dCTP, Cy5-dCTP) with fluorophor, purification and quantitative mark product
Thing.Can be used for chip hybridization with fluorophor DNA.
7) chip hybridization and cleaning：Take 1 μ LcDNA and be dissolved in hybridization solution, 45 DEG C of hybridized overnights.Chip is taken out in rich Austria Slide
Washer8 chips are washed dry instrument and are cleaned, and cleaning procedure is as follows：Washing liquid I：0.2%SDS, 2 × SSC, 42 DEG C of 120S is cleaned 2 times.
Washing liquid II：0.2%SDS, 2 × SSC, 42 DEG C of 80S is cleaned 3 times.
After the completion of cleaning procedure, centrifuge dripping is to be scanned.
8) chip scanning, data analysiss, differential gene screening：Using Agilent chip scanners (G2565CA) to cleaning
Chip afterwards is scanned, and obtains hybridizing picture.Using Agilent Feature Extraction (v10.7) softwares to hybridization
Picture is analyzed and extracts data.Then data are normalized using Agilent GeneSpring softwares and difference point
Analysis, and carry out differential gene screening with GeneSpring GX softwares.
2nd, result
Long-chain non-coding chip of expression spectrum detects ENST00000418539.1 in healthy control group and stable angina pectoris group
In detected signal value as shown in Figure 1.Chip examination finds the lncRNAs of a plurality of up-regulated and down-regulated expression.It is wherein long
Chain non-coding RNA ENST00000418539.1 shows to be expressed in patients with stable angina pectoris and significantly raises that Fc values are
1.84 again, wherein, from figure, P is less than 0.05, with statistical significance.In view of it may be in patients with stable angina pectoris
In exist specific expressed, the present invention enters the repeated authentication of row index by following examples using the sample of chip detection.
Embodiment 12：QRT-PCR repeated authentication long-chain non-coding RNA ENST00000418539.1 are in stable angina pectoris
Differential expression in patient and Healthy People.
First, experiment material
The sample of chip testing is chosen, the differential expression of long-chain non-coding RNA ENST00000418539.1 is carried out
QRT-PCR is verified.
2nd, experimental technique and result
1. primer specificity identification
(1) design of specific primer：Long-chain non-coding RNA is extracted from Ensemble data bases
ENST00000418539.1 related transcript sequence, and with the design of primers instrument for passing through NCBI according to the sequence of transcript
(Primer BLAST) designs primer；
Primer sequence after design is as follows：
Forward primer：SEQ ID No:2
Downstream primer：SEQ ID No:3
(2) by patients with stable angina pectoris and healthy human blood according to ABI companies Trizol reagents (article No. 15596-
026) reagent needed for and step extract total serum IgE, then quantitative with 7300real time PCR system nucleic acid quantification instrument
Purity and concentration that (Applied Biosystems AB) is quantitatively extracted.
Using Beijing Tiangeng biochemical technology company FastQuant cDNA the first chain synthetic agent box test kit (article No.s
KR106) the total serum IgE reverse transcription synthesis cDNA to extracting.
The first step thaws template ribonucleic acid on ice；5×gDNA Buffer、FQ-RT Primer Mix、10×Fast RT
Buffer、RNase-Free ddH2O is immediately placed on ice in (15-25 DEG C) defrosting of room temperature after defrosting.Using it is front will be every kind of molten
Liquid vortex oscillation is mixed, and brief centrifugation remains in the liquid of tube wall to collect.
Second step opens secondary structure, and reaction system and condition are as shown in table 4,
The removal system of the genomic DNA of said components is prepared into mixed liquor, is thoroughly mixed.Brief centrifugation, is placed in 42
DEG C, it is incubated 3min.It is subsequently placed in and places on ice.
3rd step reverse transcription reaction, reaction system and condition it is as shown in table 5：
By the Mix in reverse transcription reaction, in being added to the reactant liquor of gDNA removal steps, fully mix.42 DEG C, incubation
15min.95 DEG C, it is put in the cDNA on ice, obtaining after incubation 3min and can be used for subsequent experimental, or cryopreservation.
(3) long-chain non-coding RNA ENST00000418539.1 is expanded：Using the Power SYBR Green of ABI companies
PCR Master Mix fluorescence quantitative kits (article No. 4367659), by template of the cDNA of reverse transcription quantitative fluorescent PCR is carried out
Amplification.
Quantitative fluorescent PCR system is as shown in table 6；
Quantitative fluorescent PCR program：The first step 95 DEG C 10 minutes；2nd step, 95 DEG C of 15S, 60 DEG C 1 minute；3rd step, 95 DEG C
15S, 60 DEG C 1 minute, 95 DEG C of 15S, totally 40 circulation.
Electrophoresis detection, from 0120000Marker (Beijing CoWin Bioscience Co., Ltd., article No. CW0632) knots
Fruit is as shown in Figure 2：Amplified fragments size is identical with expection, amplified production only one of which band.The primer pair meets above-mentioned mark
It is accurate.The specific primer of upstream, its sequence is shown in sequence table SEQ ID No.2, and the specific primer in downstream, its sequence is shown in sequence table
SEQ ID No.2。
2. the preparation of standard DNA template
According to long-chain non-coding RNA ENST00000418539.1 base sequences (its nucleotide sequence such as sequence table SEQ ID
Shown in No.1), commission Shanghai life work synthesis.
Sampling 2ul synthetic products, are connected to Puc-TTA cloning vehicles (Beijing CoWin Bioscience Co., Ltd., goods
Number CW0801), in being subsequently transformed into DH5a competent cells.It is the special of SEQ ID No.2 and SEQ ID No.3 by sequence
Property primer screening positive colony, extract plasmid DNA, plasmid DNA is fixed with 7300real time PCR system nucleic acid quantifications instrument
Amount, and do 10 times and be serially diluted as standard curve that (standard DNA template concentration range is 102-106Copy/ul).
3. sensitivity experiments are by standard
Standard DNA template plasmid is diluted in proportion 102、103、104、105、106Copy/ul, carries out quantitative fluorescent PCR
Detection sensitivity.Concentration limit is 102Copy/ul.
4. cRNA templates are synthesized
The total serum IgE for taking the angina pectoriss of aforementioned stable type and Healthy People is each 5, using Beijing Tiangeng biochemical technology company
FastQuant cDNA the first chain synthetic agent box test kits (article No. KR 106) synthesize cDNA to the total serum IgE reverse transcription extracted.
The first step thaws template ribonucleic acid on ice；5×gDNA Buffer、FQ-RT Primer Mix、10×Fast RT
Buffer, RNase-Free ddH2O is immediately placed on ice in (15-25 DEG C) defrosting of room temperature after defrosting.Using it is front will be every kind of molten
Liquid vortex oscillation is mixed, and brief centrifugation remains in the liquid of tube wall to collect.
Second step opens secondary structure, and reaction system and condition are as shown in table 7：
The removal system of the genomic DNA of said components is prepared into mixed liquor, is thoroughly mixed.Brief centrifugation, is placed in 42
DEG C, it is incubated 3min.It is subsequently placed in and places on ice.
3rd step reverse transcription reaction, reaction system and condition it is as shown in table 8：
By the Mix in reverse transcription reaction, in being added to the reactant liquor of gDNA removal steps, fully mix.42 DEG C, incubation
15min.95 DEG C, it is put in the cDNA on ice, obtaining after incubation 3min and can be used for subsequent experimental, or cryopreservation.
5. fluorescence quantitative PCR detection long-chain non-coding RNA ENST00000418539.1
Using the Power SYBR Green PCR Master Mix fluorescence quantitative kit (article No.s of ABI companies
4367659), fluorescent quantitative PCR is carried out by template of the cDNA of reverse transcription.
Quantitative fluorescent PCR system is as shown in table 9：
Quantitative fluorescent PCR program：The first step 95 DEG C 10 minutes；2nd step, 95 DEG C of 15S, 60 DEG C 1 minute；3rd step, 95 DEG C
15S, 60 DEG C 1 minute, 95 DEG C of 15S, totally 40 circulation.
The relative table of long-chain non-coding RNA ENST00000418539.1 in qRT-PCR detection patients with stable angina pectoris
Up to amount as shown in Figure 3.According to the relative quantification formula of qRT-PCR：2-△△ct, long-chain non-coding RNA is calculated respectively
Expressions of the ENST00000418539.1 in patients with stable angina pectoris and Healthy People, as a result shown stable angina pectoris
Patient compares with the long-chain non-coding RNA ENST00000418539.1 expressions of Healthy People, and it 2-△△ctMeansigma methodss are
5.96.These results suggest that, long-chain non-coding RNA ENST00000418539.1 is universal in patients with stable angina pectoris blood
Height expression, this is consistent with 1.84 times of foregoing long-chain non-coding RNA chip results ENST00000418539.1 rise,
Wherein, from figure, P is less than 0.05, with statistical significance.Therefore, long-chain non-coding RNA ENST00000418539.1
Can be as the new blood biomarker of stable angina pectoris diagnosis, for the examination of stable angina pectoris.
Above technical characteristic constitutes embodiments of the invention, and it has stronger adaptability and implementation result, can basis
It is actually needed the non-essential technical characteristic of increase and decrease to meet the demand of different situations.
Table 1
| Reagent | Consumption |
| 5×gDNA Buffer | 2ul |
| Total serum IgE | 1-2ug |
| DEPC water | Complement to 10uL |
Table 2
| Reagent | Consumption |
| 10×Fast RT Buffer | 2ul |
| RT Enzyme Mix | 1ul |
| FQ-RT Primer Mix | 2ul |
| RNase-Free ddH2O | Supply to 10 μ l |
Table 3
| Reagent | Consumption |
| Power SYBR Green PCR Master(2×) | 5ul |
| CDNA samples | 0.5ul |
| Forward Primer(10μM) | 0.25ul |
| Reverse Primer(10μM) | 0.25ul |
| Nuclease-Free Water | 4ul |
| Total capacity | 10ul |
Table 4
| Reagent | Consumption |
| 5×gDNA Buffer | 2ul |
| Total serum IgE | 1-2ug |
| DEPC water | Complement to 10uL |
Table 5
| Reagent | Consumption |
| 10×Fast RT Buffer | 2ul |
| RT Enzyme Mix | 1ul |
| FQ-RT Primer Mix | 2ul |
| RNase-Free ddH2O | Supply to 10 μ l |
Table 6
| Reagent | Consumption |
| Power SYBR Green PCR Master(2×) | 5ul |
| CDNA samples | 0.5ul |
| Forward Primer(10μM) | 0.25ul |
| Reverse Primer(10μM) | 0.25ul |
| Nuclease-Free Water | 4ul |
| Total capacity | 10ul |
Table 7
| Reagent | Consumption |
| 5×gDNA Buffer | 2ul |
| Total serum IgE | 1-2ug |
| DEPC water | Complement to 10uL |
Table 8
| Reagent | Consumption |
| 10×Fast RT Buffer | 2ul |
| RT Enzyme Mix | 1ul |
| FQ-RT Primer Mix | 2ul |
| RNase-Free ddH2O | Supply to 10 μ l |
Table 9
| Reagent | Consumption |
| Power SYBR Green PCR Master(2×) | 5ul |
| CDNA samples | 0.5ul |
| Forward Primer(10μM) | 0.25ul |
| Reverse Primer(10μM) | 0.25ul |
| Nuclease-Free Water | 4ul |
| Total capacity | 10ul |
Sequence table
<110>No.1 Hospital Attached to Xinjiang Medical Univ.
<120>ENST00000418539.1 and preparation or diagnostic agent or medicine or test kit and application
<130>
<160>
<170>
<210> 1
<211> 13458
<212> DNA
<213> Homo sapiens
<400> 1
GAGGATCATATTTTGAGCCCATCACACATTCATGCATGCCATGCCATTTTTCTTAGTTTGTTGTTTCATTCAT
TCATTCATGTATTCATCAATACCTTCCACAGATACCTATTGATCTGTTTTTACAGGCCCATGTCACTATTCTCAGGT
TGTCAGTGGCTATCTGTGTGACCCCCACCTCCCAATCCCATACCTTCTAGAGCAGCAGTCCCAACCTTTTAGGCAAC
AGGGACCAGTTTCATGAAAGACCATTTTTCCACAGACTAGGGCGGCAGGGGATAGTTTAGGGATGATTCAAGTACAT
TACATTTATTGTGCACTTTATTTCTATTATTATTACTTTGTAATATATAATGAAATAATTATACAACTCACCATAAG
GCAAAATCAGTGAGAGCCTTGAACTTGTTTTCTTGCAACTAGGCGATCCCATCTCGGAGTGATGGGAGACAGTGACA
GATCATCAAGCATTAGATTCTCACAAGGAGAGTGCAACCTAGATCTCTGCCATGCGCAGTTCACTAGAGGGTTCACG
CTCCTATGACAATCAAGGTCGCCACTGATCTGACAGGAGGCCAAGCTCAGGCAATAATGCTAGTGATGGGGAGTGGC
TGTAAATACAGATGAAGCTTGATTTGCTTGCCTGCCACTCACCTCCTGCTGTGCGGCCTGGCTCCTAATAGGCCAGT
ACTGGTCTGTGGCCCAAGGGTTGGGGACCCCTGCCCTAGAAGAGTTCTTCATCTGTACTCATGAAAAGTTGATAAGT
CACCTAAATCACCTTTAATAAGAGCCCAGTAAGTTGAGATGTCACAAAAACAGAGCCAGAAAAAGAATCTATACCAT
CTGCTACAGCCAGGACCAGCTACCTAATTTACAAGTCTCAGCACAAAATGAAAATGTGGGGCCTCAGCCAGATGTGG
TGGCTCATGCCTGTTATCCCAGCACTTTAGGAGACCAAGGTGGGAAGATTGCTCGGGCCCAAGAGGTTGAGGCTGCA
GTGAGCCGTGATTGCGCCACTGTACTCCAGCCTGGGCAATAAAGCAAGACCCTGAAAAAAAAAAAGAAGAAAAGAAA
AGATGGGTAGGGAAAAAAGTGCCATTAGAGGCACTAAACTACAAACTTTTCCTTTCTTCTGTGGTCTCTGTCTTGCC
CTCTCTTGGTGTTTCTTATTTACTGTTTGATGCTATGCTTCCTTGGGCACAGGGATACTTACAGGGTGAGTGCAGAC
CCTCACAGGCACAGGGAGGGTGAGGAGTAGGAGGTGGGCTGCCCGTGACTGCATATCCATGCACAGTTCACCAGCTC
CAGGAGTCTTCCTCTGGCTAGACCACCAGACTGACATGCCATATCTGCCCAGGGGCAAGGATGAGGTGAGGCCGTCA
ATCTTCCCTTCCCACAGGCCTGCTGCCCCAACCCATGGCTGCTGGGAGACCTCCAGGGGATTAAATCTCCATTCCAA
AGCAGTCTCAGCACCTGGCTAGGGGATGGCTGAGAGGCCAGTCCTGCTGCTGCTCATCAAAGGCACCATGGCGCTGC
CAGCCCAGGGCAAGGATGGCTGCCACCCTGCCTCTCCCCAAAATGCCATAGGGCACACCAGCTCAACCCCAATTCTC
CCTGTGCCCACAGCTAGGCCCTTACAGCAGGCAGAGGTCAGCAAGGAAGGGGAAGTCAGGTGGGGCCTGGGGAACCA
GGAAGCGGGGAACAGGCAGCTGAGAATTGTTCCAGGGAAGCAGGGAGAGGCAGACCACGCATGGGCCAAGACTCCAA
GCCCTCAGTGCATGCTGCATTGTCCCATTGGACTTCACTTGCAAAGTGTAAATGCAACGAAAAAATTATTAAGAATT
TCAAGACAAGGTGGGTGCAGTGGCACTTACCTGTAGTCCCAGCTACTCAGGAGGCTGAGGCGGGGGAATCACTTGAG
CCCAGGAATTTGAGTCCAGCCTGGGAAGCATAGCGAGACCTCCCACCTCAAAAAAACAGAAAAGAATTCTAAGACAG
TGTCATAGAACATTAGCCCCCCAGTGCAGGACCCTTTTGAACAGAGCTCTTCTGAGCACGAGGCCCTGTCTATGCCT
GATGCACACCCACGAAACCGACCTTGGCTCCAGCTTTCAATGCGAACTTCTTACAGTTTAGGTCCTCTGCGAAGTCA
CTTGCTTCCAGTAGCTCCTATCAGAGCTGAGCTGAGGCCTTAGACTGGGGTTAAAAGAGAGGTGAAGTTTCCCGCTG
CAGCACTAAACACATCACTGAAGGCCTACGGAGTAATCCTTGAGCATCCCCTGACTCAGAGCACTGCCACCGCATCA
GCTTCTTCCAGATGCCAAGAAGCACCTCTGGGTCTTGGGGTTCCGGCTTGTTTGTGTCACAAGTAATAACTTGTATT
TTATTTGCTTTTAAAAACTGGATATGCTCCCACTTCTCAGCCTCTACAATGAGACCTGTGTTTCGTGTTTTTCAGTG
TGCCTTTGAGAAAGGCCCATAAACCTATAAACACCCTTTTTCTCCCCAAGGTGAGGTTTGACCCTAGTACTTACTAC
CACTGTTTACTGGAGAGTCATGTTGAGAACATCTCCTTTAGGCTCTATTTCTCATCCTAAGGGATATATCCAGCCTT
CATTATGGAAGGTTTTGAAGGCCTCAAATAAAATCAAAGTAGAATTTGAATAAAACATCTTGGCTGGGCACGGTGGC
TCGCATCTGTAATCCCAGCACTTTGGGAGGCTGAGGTGGGCGGGTCACAAGGTCAGGAGATCAAGAACATCCTGGCT
AACACGGTGAAACCCCCATCTCTGCTAAAAATACAAAAATCAGCTGGGCGTGGTGGCGCGTGCCTGTAGTCCCAGCT
ACTCGGGAGGCTGAGGCAGGAGAATCGCTTGAACCAGGGAGTCGGAGGTTGCAGTGAACCGAGATCACACCACTGCA
CTCCAGCCTGGCGACAGAGAGACTCTGTCTCAAATAAATAAATAAAACATCTTAAAGAAATAGAAAATACTAGTAGA
GTATTCACTCTGATTTCAGAATCAATTTGTGGAAATTCAGTTTCACAAGAATGAAAAAAGCCCCCTAAGCAAGGTAG
CTTTTTTTTTTTTTTTTAAAGATGGGGTCTCACTATGTTTTCCAGACTGATCTCAAACTCCTGGGCTCAAGCAATCC
TCCTCCCTCAGCCTCCTGAGTGGCTGGGACTACAGGTACACATCACCACCACCCTGCTAGGATATGGTAGCTTTTTA
ACCAAAACACACCACCATGCAATGGAGAGCATGTAACTTGTTATTTGGTCATACAAATACATCAGAATAAATTCCAC
CTCTCTTGTGGAGCCCTTTTTCCTACATCTGCACAGAAACTGGCACTAGCAGGACTGCAACCGAGGCAGACTCGAGA
ATTAGCAGACTCTGGCTCAGGCAGGACAATAGAGCAGCGGGAAATTTCTTTCAAGGGCTTTTGTTGCTGTTGTTGAG
ATGGAATCTTGCTGTGTCACCCAGGCTGGAGTGCAGTGGTGCTGTCTCGGCTCACTGCAATCTCCGCCTCCCAGGTT
TAAACGATTCTCCTGCCTCAGCCTCTCAAGTAGCTGCGATTACAGGCATGTGCCACCATGCCCAGCTAATTTTTGCA
TTTTTGGTAGAGATGGGGTTTCACTATGTTGCTCAGGGAGGTCTCGAAACTCCTGACCTCAGGTGATCCGCCCACCT
CGGCCTCCCAAAGTGTTGAGATTACAGGCATGAGCCACTGCGCCCGGCTTGAGACCTTGAGGGGCTTTTTAACCAAG
AGATGGTGTAGGGACAGAAGCCAGCAAGAAGAACATGGTGGACTCTCTGTGAGGCCCCAAACCCCAAATCATGAGCC
TCCGCCCCCTGAAAGTATTCCAGGATCTCTGAGGGAGGTTTTGGGAGGGGGCCAAGGTCCTGCAATTGGGTGGGTGG
GGCTCCAAGCCATTGTTCTTTGCACATTCATGCCAATGTGACCTAATTGGCTAAGCACAGGCTGGAGAAGCTAGATA
AACCTATTTAAATTCAGAGCCAAAAGACACTCAAGGATGTTTTCTGAGGGGGTGTGAGGGTTGGAAAATCATCCTAT
ATTTGATTTATCTCCCACCTCATGTCCTGAGTCAATTCCTTAGTCTTAGATGAGGGAGACGACTTGGGAGTCATCCT
TGTTTTTGATGAGCTATATAACCCTATGGCCAGCAGAGGGAAGTACTGCATTTCAGAGCGACAATTTGAGATCTATG
AAAGAATTTCAATCGAGAATAAGAGGCCGGGCGCGGTGGCTCACGCCTGTAATCCCAGCTCTCAGGGAGGCTAAGAG
GCGGGAGGATAGCTTGAGCCCAGGAGTTCGAGACCTGCCTGGGCAATATAGCGAGACCCCGTTCTCCAGAAAAAGGA
AAAAAAAAAACAAAAGACAAAAAAAAAATAAGCGTAACTTCCCTCAAAGCAACAACCCCCCCCCCCCTTTTCATATT
CTTGCCTGGAAGAAAGGCCTTGCTTTCCTCAGCTTCTTAAAGCTGGGAGAAGTTAAAGCCATTCTGAAATGCTGCTG
CTACTGCTATATGTGGGGGAGAAAGCAAATTTTCTTTTTACTTTTTTAATCAGTATTTATTTTAAGTTCGAGGGTAC
ATGTGCAGGATGTGCAGGTTTGTTACATAGGTAAACGTGTGCCATGGTGGTTTGCTGCACAGGTCAACCCATCACCT
AGGTATTAAACCCAGCATCCATTAGCTATTCTTCCTGATGCTGTCCCTCCTCCCACCCCACCAACAGGACCCCAAAT
TTTCTTTCTTTTTTTTTTTTTTTTTTTTTTGAGATGGAGTCTAGCTCTGTCACCCAGTCTGGAATGTAATAGCTCAA
TCTCGGCTCACTGTAACCTCCATCTCCCAGGTTCAAGTGATTCTCCTGCCTCAGCCTTCCAAGTAACTGGGATTACA
GGCATGCACTACCATGCCTGGCTAATTTTTGTATTTTTAGTAGAGACGGGGTTTCACCATGTTGGCCAGGCTGGTCT
CAAACTCCTGACCTCAAATGATCCGCCTGCCTCAGCCTCCCAAAGTACTGGGATTACAGCTGTGAGCCACCGTGCCC
AGCCTCCAAATTTTCTTATTTCCATGGCCTTGAGCAAAATTATACCATCCTAGACAAACACACACACACACACACAC
ACACACCATTCTCCCCCTCTTGCTGTCTTTCTCAGATATGAGTCTCTCCTCCAATGGCCATTTTAAGCTTGCTCTTT
TGCTCTGTAATACAAGGCCTGTTCCCTGTTAAAAGGAATGTCAGGCATCTGTGAGCAGGTAGGGAAAACATTGCAGG
AGGCAAGTAAACTTCTCAGCATACCTGCCATCATTCATACCTTGAAGGCAGGTCAAGCCTGTAGCGTCTGAGTCTTC
CATCCCTGAAGCACCAGCAGCTCTGACTAGAGGTCCTGTCTCCCAGCTGTGGCCCCAGCTCTCCCTGATGGTGACAG
TTGCGACTTCCAAACTTGCTCCTCCCCATATCTGTAACTGAGCCCAGCGTACATTTGCCTCATGGGTGCCATTGGAC
TTCCTTTGTGAGCCATGTTTTCTTTCACCTTGTCCCAATCCACAGTTAACCAAATCTTGGCCCAAATTATGTGCTGG
CTGGTGTTGGGCAGCTGTGAGACCCCTCCTTGCCCCATCTCTGTCTTCTAGGCTCTTCTGAGAACCAGTGTCCTTTG
AGATCTCAGATGGGATTTAGAGGAGGGTTTCCACCCAGACCACTTTCCACAAATTTCACTTCTGCATTTCATCTGGT
AGGGTGTGGTTTTCAACTAGAAGCCCAAATGAGGAAGAAAAAAATTCATTCACCATTCAGGACAGGGTGTTCACATT
TGAACCAGGCAGAAATAGCTGAAAGAAACTGAATTCTAAAGGGACTCTCAGCTTTTGGGGGAGACATATTTGAGTGC
AGGAGGCTAGACCACCCACTGGCTGGCTCTTTGCTCCTCACTGAGGCCAGCCTTGACTCTAAACAGATAACACAGGG
CCAAAACACACCCAGCCAGCTTTGATAAGCAACCGTCTCTGCCAAACAGTTTCTTTCACAGGGCCCAGATGATCTCA
TCACAGAGGCTGCATTCTCTGGTATCTGTTCCAAGCACAGACTCACTTCTCATTACCTTTGCAAATAATAGTCAATC
TATCCTGAGAAGCTCCTTTATATCCCATACTGGGCTAAGGTCTGAAGAGGGTACAGTTAAAGAACCCATGATCTCAG
ACTGGGCATGGCAGCTCATGCCTATAATCCCAGCACTTTTGGGAGGCCAAGGCAGGAGGACCGCTTTAGCCCAGGAG
TCCAAGACCAGCCTGGGCAACATAGTGAGAACTCTTGAGACCTTGTCTCTACAAAAAATAAACAAAATTAGCCATGC
ATGGTGGCTTGCACCTGCCTGTAGGAAGCTGAAGCAGGTGAGCCTGGGACGCAAAGGTTGCAGTGAGCCAAAATCGC
ACCACTGCACTCCAGCCTGCGCGACAGAGACTGTCTCACAAAAAAGAAAAGAAAAGAAAAAAAAAAAAGGACCCATG
ATTCCTCATGAAGCTTGCCTCTGGATCTAATTCATGGTTCTCTATTTCCACAAGCTTTGTGGAGCAAGGGGCTCATG
ACACTTCATCTTTATGGTACAGATAACTCACCGAAACTGGATTCCTTTAGCTACTGGTCCCACACCACTCCCTCCAG
GAGCCACCTTCCCTCTCCATGTTTTTCTCATGAATTTTGAACCGGCCATTTCTATTTTATTTTATTTTACATTTATT
TATTTATTTATGGAGACAGAGTTTCACTCTTGTCACCCAGGCTGGAGTGCAGTGATGCTATCTTGGCTCACTCCAAC
CTCCACCTCCCAGGTTCAAGTGGTTCTCCTGCCTCAGCCTCCCAAGTAGCTGGGAGTACAGGCACGTGCCACCACAC
TCGGCTAATTTTTGTATTTTTAGTAGAGACGGGATTTCTCCATGTTGGCCAGGCTGATCTCGAACTCCTGACCTCAG
GTGATCCACCCGTCTCGGCCTCCTAAAGTGCTAGGATTACAGGTGTGAGCCACTGCGCCCAGCCAAAATAAAAATAG
ATTTTAAAAATGAAAATAAAAATAACATAATAAGATGAACACCACCTCGGTGATCTGTGGGATAGTATCAAGTGGTC
TAACATAATTGGAGCCCCCCCAAAATGAGGGAAGGCTAAAAAAAAAAAGCCAAAATTTTTCTAAACTTGAAACTGTT
TAAAGCAAAAAATAATAACAATGCCTTATGAGGTTTATAACATATGTAGAAATAAAATGAATAATCATAATAGCCCA
TCAGGGGGCTGAGGTGGGAGGATTGCCTGAGCCCAGGAGTTTAAGGCTGTGATGCACTATGATCATGCCTGTGAATG
GCCTCTGCACTCCAGCCTGGACAACATAGTGAGACTCTGTCTCCAAAGAAAAAATGGAGGAAAAAAGAAAAAGAAAA
AGAAAGAAAATAATGGCCCAAAGGATAGAAGAGAAGAAATGAGAATAGACTGTTATAAAGCTCTTCGATTTTGTGTG
GAGGAGTATAATAACATTTGAAGGTATTCTGGGTTAAGCAGTTTATCCGAGGTGTCACATATGATGGAGCTAGGATC
CAAATTAAGCTTTCCAGATTCAAAGCTTTTTGTGGGGAGAGGAGATGTGAGGTTAGGCTCACACATGCCACAAGACA
GTTTCCCCAAAGGCTGCGGGAATTAAGTGCTTTCATCTCTGTAGTGGGTATTCCCTCAAAAGACGTGTTCAAGTCCT
AACCTCCATACCCAAGAGAGTCAGCTTACTTGGAAATAGGCTCTTTGCAGACGTAATCAAGTCATACTGAGTTAGGG
TAAAGCCTAACCCAATGACTGGTGTCCTTATTATAAAGGGGGGGGTGCGGTACCAGGTGTGGTGGCTCACATCTGTA
ATCCCAACACTTTGGGAGGCCAAGGCCAGTGGATCACCTGAGGTCAGGAGTTCAAGACCAGCCTGGCCAACATGGTG
AAACCCTGTCTCTACTAAAAATACAAAAATTAGCCGGGCATGGTGGCAGGCACCTGTAGTCCCAGCTACTCGGGAGG
CTGAGGCAGGAGAATCACCGGAGCCCGGGAGACAGAGATTGCAGTTAGCCGAGATTGCACCATTGCACTCCAGCCTG
GGTGACAGAGCAAGACTCCATCTCAAGAAAAAAAAAAAAAAAAAAAAGGACATTTGAACACAGAGAAGACACACAGG
GAAAATGCCATGCAATGATAGATATCTATAAGCTGAGGAACACGCCCAGAAGTAAGAAAGAGGCAAGGAAGGCTCTC
TGAGAGCCTTCAGAGAGTGTGAGGCCCTGCCCAACACCTAGATTTTGGACTTCTGACCTTCAGAACCAGAGAGAATA
TATTTCTGGGTTTTGTTTGTTTTTTTGTTTTTCTGAGATGGAGTTTTGCTCTTGTTGCCCAGGCTGGAGTGCAGTGG
CATGATTTTGGCTCACCGCAATCTCCACCTCCCAGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCAAGTAGCTGGGA
TTACAGGCGCCCGCCACCATGCCCGGCTAATTTTTTCGTATTTTCAGTAGAGATGGGGTTTCTCCATGTTGGTCAGG
CTGGTCTCGAACTCCTGACCTCAGGTGATCCTCCCACCTCAGCCTCCCAAACTGCTAGGATTACAGGCATGAGCCAC
CATGCCCGGCCATATTTCTGTTGTTTTAAGCCACCTTGTCTGTGGCATTTTGTTACAGCAGCCCTAGGGAACTAATA
CAATCTCTAAAACTTCCTCTTTTTGATGCTCCCAGTCTTTAGAATGTAAAGAAGCTCCGCCCCTACCCATGCATGGG
CCAAGTATAAAGTGAGTGTTGATAGCAGGATATTAAGTTGACAAAATAAACCAGATTCAGGATCTAAGAGCCACAGG
GGAAAATCATGAAGTGGGGACTGGTTGGACTGGTTTTTCTCATAAATCTCAATGACTTTGATCCCAACCAGGTGGAA
TTTTAAAGTTAATTGATTTAGGGAGGAACACCTGAGCAGCTGCACCGTGTGTGAGCCCTGTGCTAGGCCCAGTGGGG
AAGCCAGGTGAGTCAGGTGCTGCTGCCTTACGGGAGCTCATAGGATTCGCCTCTACAGAAGGTGTGATGGAACAGGT
CAGTCACTAGCCAAGGTGTGAGTAACAGGCTGTGCAAGTTCAAAATCAAGAGAGTGATTCATTTCCCCTGGGGGAGC
ATCAGCCAGACTGCTTCACTGAGGAGAAGGCAGTGGCGACATCTCTCTAAATATCTGACTGTGGAATCCATAGCCCA
GAGAAGGTCTTGTGTGTCGATTAGAGACAAGCAAAGGAAAACAAATCTACCAGGAATGCTGGTGTTTTCCTGGAGAG
TGTTTCACTGATCACGGGGTAGTGCTCTTTGCTCAGTGGAGAGGTACAAAGCTGTCTAGCCCCTGCTTGCTCTGAGG
GGGAATTCTCTGGGCTCTCGGAACCTCTTATTTTCCCTTCTGGTTCTCCACTTGGGGTGAGTGCTTATCACCACCTT
CTGTAGAGATTGAGGAAAGTGGAGAAGGTACACAGAGGCTTTGGAGTTGCTAGCGATCTCAATGACATCTTGTAAGA
GAGCTGGTGGCTCAAGAAGGAGAGAGCAGGCCCTTGTGTTGCATTTGCAGGATCTGGGGCAATAGTACAGCCCACAT
ATCTAAGCTCCATTTACGTTGCCAAAAATAGCATAGGCCACACATCTAAGCTCCATCTACATTGACACTCCATGACC
CCACGTGGGCCACCCCAGTGTCATGGCCCAGTCTGGAGAAGTGACCCTGCGAAAAAGCTGTGTATGCCCTGTAGGTA
GACTTGACCCTTCTGTGCAAGAAATTTTGATGTCCTGCGAGCACCCAGAATGGTCTAGGAGAGAGGGAACAGGCTCC
AGTGGACATATTCTCTTCGCCCTGTTGACTCCCTCCTCCATGGGAAGAGACTTGGCTTAAACAAGCCTAGAGCAGGT
CTGTTTAAGGTCTGGGGCCCTGAGTAGGAATCTACTTACCCACGTCGAAGGTAGAATTGGGAAGAAGCACTGTTATA
AACTCAACAGAAAGTGATACTGAGTTTCATTTTTTTTTTTTTTTTTTTTTTTGGAGACAGTCTTGCTCAGTCGCCCA
AGCTGGAGTGCAGTGGTGTGATCTCGGCTCACTGCAAGCTCCGCCTCCTGGGTTCACGCCATTCTCCTGCCTCAGCC
TCCTGAGTAGCTGGGACTACAGGCGCCCACCACCACGCCTGGCTAATTTTTGTGTTTTTAGTAGAGACGGGGTTTCA
CCATGTTGGCCAGGATGGTCTCTATCTCCTGACCTCGTGATCCGCCCGCCTCGGCCTCCCAAAATGCTGAGATTACA
GGTGTGAGCCACTGCACCTGGCCAATACTGAGTTTCTTGGTTTTGCTCTGGAACTAGACTAATGAATTCCAATCCTC
ATGGAAGCCCTGTGTGAAGCATTTCATGGAGAAAAATTTGTATGATGCAGTGGTTTGCAAATGGGGGAAATTTGGCA
TACAGTGACATTTGGTGATGTCTATAGGCATTTTGCATTGTCATAATTGGAGGATACTGCAAGACGTCTGGTGGGTA
GAGGCCAGGGATATTGCAAAATATCCTACTATGTTGCCCTGCAGGATAGCACCCCATAAGAAACAATGATTTTGGCC
GGGCATGGTGGCGCACGCCTGTAATTCCAACAGTTTGGGAGGCCGAGGAGGGCAGATCACTTGAGGTTAGGAGTTTG
AAACCAGCCTGGACAACATGGTGAAACCCCATCTCTACTAAAAATACAAATAAATTAGCCGGGTGTGGTGGCAGGCA
CCTGTAATCCCAGCTACTCTGGAGGCTGCGGCAGGAGAATTGCTTGAACGGGGAGGTGGAGGTTGCAGTAAGCCGAG
ATCGTGCCACTGTACTCTACCCTGGGCAACAGAGCAAGACTCCGTCTCTAAATAAATAAATAAATAAAAATAAAATA
AAATAAAAGAAACAATGATTTGGCCCCAAATATCAATAGTGCTGAGGTTTGAGAAATGAATTTTTAGTACCCTTAGC
ACATGGTTTGGACTTGTTTGCTGCAGGATGCTGGGCCTGGAAGATATAAGATTGCAAGTAGCCACGGCTCCTAAAGA
ATGACTGAGTGGCTGCTTCCAGAAAAGTAGGTCAGAATCAGCAGAAAAACAACTTAATGAGGCCGGGCGCGGTGGCT
CACGCCTGTAATCCCAGCACTTTGGGAGGTCGAGGCGGGTGGATCACGAGGTCAGGAGTTCGAGACCAGCCTGGCCA
ACATGGTGAAACCCCGTCTCTACTAAAAACACAAAAATTAGCCAGGCGTGGTGGTGGGCGCCTGTAGTCCCAGCTAC
TCGGGAGGCTGAGGCAGAAGAATTGCTTGAACCCCGGAGGCGGAGATAGATGGCAGTGAGCCAAGACTGTGCCACTG
CACTCCAGCCTGGGCAACAGAGCACGACTCCATGTCAAAACAAAACAAAAAAAAAATTAATGGTGTACTTGGGACTA
TACTATCAGCCTCAACAAATAGACTTGGGGCCCAGCAGCTGTGCCAAGCTCCCGAAGACCCACCAGCCTGAACTGTC
CATCCCTTTTCCTCATGCAACTTTGCCACCCACATCACCCACCATCCCTCACCTTTGGTCTTCACTGGGGCAGCTCT
CCATTCAACTAACTCCCCCGCGCTTAAGTTAATTATCTGAGTTTTCTTTGCAGCTTAATACTTACCAGGCCTCATGG
TGTCTACTCTTCTTTTTTCTTTTTTTTCCCAGACAGAGTTTTGCTCTTATTGCCCAGGCTGGGGTGCAATGGTGCCA
TCTCGGCTCACTGCAACCTCCGCCTCCCAGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGAATTATA
GGCATGTGCCACCACGCCCGGGTAATTATGTATTTTTAGTAGAGAAGGGGTTTCTTTCATGTTGGTCAGGCTGGTCT
CAAACTCCCAACCTCAGGTGATCAACCTGCCTCAGCCTCCCACAGTACTGGGATTACAGAAATGAGCCATTGCACCT
GGCCCAGTGTCTACTTTTCTTTATCAGGCTAAACAGCTCAGGACACAGATAGAATTTGGACTTAGGTCCGTGTGAAT
CCAAAGGGCCATGCTCAACTGAGCAGCCGGCACTGCCTCGCTGAACAGCCAGGATTCTGGGAGTGCTACTTTGTACA
ACATTTTATGAAATGTTTCTGGCCTCCTTCTAGACCAGAGCAGAAAGGGGTGGTTAGGATGTTCTAAATACCAACCC
CCTCTACTCCATCCCCAGGTTCATGACAACTTTTTAAGCTGAGCATACTGCAATCACTTAATTCTTCTAGACCTAAG
ATAAGCCAGTAGCCACGTCCACTCTGAGCCTGTGAGCAGGCAGGATCAAGTCTAAGAGCATCTCTTCTTTGGAATCT
TAAGGCCTAGGATCCCAGCAGTTTTGGGGTCTAAAGACTTTTCAGGCTCCCGAAACTCCCTCTATTTTTAATAATTT
TAAATGTTTCCCCAATACTTAGCTAAGCAAAGAAGGAAAAAGTGTACTTTTATTACACAGTCAGTTCTGGATAATTC
TAGGGGAAATTCTTCCATTTTAGAAATCTCTGCTTTCTTCTTTTCTCCCAGATTCTTCTCATTAATTCCCTCAGTAA
ATATTTGTCTGTGGCAGGGTGCAGTGCTCGTGCCTGTAATCCCAGCACTTTTGGAGGCTGAGGTGGACGAATCACCT
GAGGTCAGGAGTTTGAAACCAGCCTGGCCATCATGGTGAAACCTCGTCTCTACTAACAGCACAAAAATTAGCTGGGC
ATGGTGGTGCGTGCCTGTAATCCCAGCTACTTGGGAGGCTGAGGCAGGAGAATTGCTTGAACCAGGAGGCGGAGGTT
GCAGTGAGCCAAGATCACGCCACTGCATGCCAGCCTGGGTGACAGAGCAAGACTCTGTCTCGGGAAAAAAAAAAAAA
AAAAACTATATATACATACATACATATATATATATATATATATAAAATGTACATATATATTTATCTGGCACCTACTA
GATGCCCCAGGTACCATGGTAGGCACAGGTGTACAGTGATGAAGAGCATTTTCCTATAAAATATTACCCAGGCTGAC
TCCAAGTTTTAATACCTCAGGTAGGAAAAGGAGGAGAGTGTTAGTAAAATGTTTGCCGTTGAATTTCACCAGCGTAA
AAACAACATTCTGGGAAATAATTATCTGCACTTCAGTTCTTTGGGGTGGTACCGGAGGGAATCTTTACAAAGGTGGT
ATTAATGGAGAAGAGGAAGGGAAGGATCCTTAGTGATGGGTTGTTTTTCCAAGCACCTCGATCTAGTGGATTTCCTT
TTTTTTTTTTTGAGATGGAGTTTCGTTTTTGTCGCCCAGGCTGGAGTGCAATGGCGCGATCTCG
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
<400> 2
ggtggtaccg gagggaatct
<210> 3
<211> 21
<212> DNA
<213>Artificial sequence
<400> 3
ccactagatc gaggtgcttgg
Claims (10)
1. a kind of long-chain non-coding RNA ENST00000418539.1 of expression high in patients with stable angina pectoris blood, its
It is characterised by the sequence such as SEQ ID NO of long-chain non-coding RNA ENST00000418539.1:Shown in 1.
2. the long-chain non-coding of expression high in patients with stable angina pectoris blood according to claim 1
RNAENST00000418539.1, it is characterised in that long-chain non-coding RNA ENST00000418539.1 is
ENST00000418539.1:1 total length or fragment.
3. long-chain non-coding RNA ENST00000418539.1 according to claim 1 and 2 in a kind of vitro detection blood
Preparation or diagnostic agent or medicine or test kit, it is characterised in that said preparation or diagnostic agent or medicine or test kit include specificity
Primer pair, standard DNA template, PCR reactant liquors, wherein, specific primer is to including forward primer and downstream primer, forward primer
Nucleotides sequence be classified as SEQ ID No:2, the nucleotides sequence of downstream primer is classified as SEQ ID No:3.
4. the preparation or diagnostic agent or medicine of long-chain non-coding RNA ENST00000418539.1 according to claim 3
Or test kit, it is characterised in that the test kit is fluorescent quantificationally PCR detecting kit.
5. the preparation or diagnostic agent or medicine of the long-chain non-coding RNA ENST00000418539.1 according to claim 3 or 4
Thing or test kit, it is characterised in that specific primer for SYBR Green, Taqman probes, molecular beacon, double cross to visiting
The detection of pin, combined probe.
6. the preparation or diagnostic agent of the long-chain non-coding RNA ENST00000418539.1 according to claim 3 or 4 or 5
Or medicine or test kit, it is characterised in that PCR reactant liquors are fluorescence quantitative PCR reaction solution.
7. the preparation or diagnostic agent or medicine of long-chain non-coding RNA ENST00000418539.1 according to claim 6
Or test kit, it is characterised in that fluorescence quantitative PCR reaction solution includes dNTP, Mg2+, Taq enzyme.
8. the preparation of the long-chain non-coding RNA ENST00000418539.1 according to claim 3 or 4 or 5 or 6 or 7 or
Diagnostic agent or medicine or test kit, it is characterised in that also including fluorescent dye.
9. the method for a kind of detection long-chain non-coding RNA ENST00000418539.1 according to claim 1 and 2, its
It is characterised by carrying out in the steps below：The first step, extracts blood total serum IgE；Second step, prepares cDNA；3rd step, quantitative amplification is long
Chain non-coding RNA ENST00000418539.1.
10. a kind of long-chain non-coding RNA ENST00000418539.1 according to claim 1 and 2 is being prepared for examining
Survey the test kit of stable angina pectoris or the application of the novel targets as detection stable angina pectoris medicine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611123473.6A CN106676109A (en) | 2016-12-08 | 2016-12-08 | ENST00000418539.1, preparation or diagnostic agent or medicine or kit, and application of ENST00000418539.1 |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611123473.6A CN106676109A (en) | 2016-12-08 | 2016-12-08 | ENST00000418539.1, preparation or diagnostic agent or medicine or kit, and application of ENST00000418539.1 |
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| CN106676109A true CN106676109A (en) | 2017-05-17 |
Family
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| Application Number | Title | Priority Date | Filing Date |
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| CN201611123473.6A Pending CN106676109A (en) | 2016-12-08 | 2016-12-08 | ENST00000418539.1, preparation or diagnostic agent or medicine or kit, and application of ENST00000418539.1 |
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| CN (1) | CN106676109A (en) |
Cited By (1)
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| CN111696627A (en) * | 2020-03-26 | 2020-09-22 | 上海生物芯片有限公司 | Design method of long-chain RNA specific probe |
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