CN110095611A - For the molecular marker TM4SF19 of diagnosing atherosclerotic and application - Google Patents

Abstract

The present invention provides a kind of for the molecular marker TM4SF19 of diagnosing atherosclerotic disease and its application, the application in reagent for disease diagnosis caused by atherosclerosis, in-vitro diagnosis and/or risk stratification of the diagnostic reagent for disease caused by atherosclerosis are being prepared or screened including providing cross-film -4-L6 family protein 19 (i.e. TM4SF19 protein) or its peptide fragment as biomarker.Present invention discover that can predict the generation of disease caused by atherosclerosis by the TM4SF19 protein concentration in detection patient tissue samples, or above-mentioned disease is diagnosed, risk stratification etc..

Description

For the molecular marker TM4SF19 of diagnosing atherosclerotic and application

Technical field

The present invention relates to technical field of biological, more particularly to the molecular marker for diagnosing atherosclerotic TM4SF19 and application.

Background technique

Atherosclerosis (i.e. AS, atherosclerosis) is one kind using disorders of lipid metabolism such as cholesterol as lesion The chronic complex disease on basis, its main feature is that arterial disease is since inner membrance, basic lesion include endarterium lipidosis, Inner membrance stove shape fibrosis, monckeberg's arteriosclerosis etc., thicken, lumen of vessels is narrow so as to cause arterial wall hardening.Lesion is often involved greatly Middle artery, is enough to block lumen of artery once developing to, then the tissue or organ that the artery is supplied can cause ischemic or necrosis The complication such as coronary heart disease, cerebral infarction, acute coronary syndrome.

At present worldwide, cardiovascular disease has become one of disease incidence and the highest chronic disease of case fatality rate. " Chinese cardiovascular disease report 2017 " points out that it is the first that China's cardiovascular death accounts for the total cause of death of urban and rural residents, and AS is exactly The main reason for causing all kinds of cardiovascular diseases.Atherosclerosis initial stage and non-evident sympton, the state of an illness add over time Again, the early diagnosis and prevention and treatment once morbidity can cause life danger, therefore to atherosclerosis are particularly important.

Atheromatosis is because of studied many years, a large number of studies show that Dyslipidemia, especially hypercholesteremia Disease is the important risk factor for promoting incidence of atherosclerosis.In the case where hyperlipidemia and endothelial injuries, endothelial cell Lower gap increases, and excessive cholesterol deposition is under arterial endothelium in blood plasma, oxidized low-density lipoprotein (oxidized low Density lipoprotein, Ox-LDL) intrusion subintima.Monocyte chemotactic factor (monocyte simultaneously Chemotactic protein 1, MCP-1) secretion increases, and promote the monocyte to dissociate in blood to invading under inner membrance, and And intravascular smooth muscle cell is also to migrating under inner membrance.Monocyte and smooth muscle cell are divided into macrophage and absorb oxidation Type low-density lipoprotein, is eventually converted into foam cells.Foam cells is constantly accumulated, and fatty streaks or even Lipid Plaque are formed, It is final to promote atherosclerotic process.The early stage of foam cells occurs, and is the early sign that atherosclerotic plaque is formed. Therefore, influence of the oxidized low-density lipoprotein to foam wanshing is inquired into, there is weight to prevention and treatment atherogenesis Want meaning.

There are no a kind of Novel markers for atherosclerosis diagnosis of maturation for this field at present, therefore compel to be essential Developing can effectively prevent or the marker of diagnosing atherosclerotic.

Summary of the invention

In view of the foregoing deficiencies of prior art, the purpose of the present invention is to provide for diagnosing atherosclerotic Molecular marker TM4SF19 and its application lack the marker etc. to atherosclerosis diagnosis for solving in the prior art Problem.

In order to achieve the above objects and other related objects, the present invention provides 19 (i.e. TM4SF19 of cross-film -4-L6 family protein Protein) or its peptide fragment as biomarker preparing or screening answering in reagent for disease diagnosis caused by atherosclerosis With the diagnostic reagent is used for the in-vitro diagnosis and/or risk stratification of disease caused by atherosclerosis.

Optionally, disease caused by the pulse atherosclerosis includes cardiovascular disease.

Optionally, the cardiovascular disease includes chronic cardiovascular disease.

Optionally, the chronic cardiovascular disease includes coronary heart disease.

Optionally, the coronary heart disease includes chronic coronary, unstable angina, myocardial infarction, Stable angina pectorsis.

Optionally, the TM4SF19 protein level of Disease caused by atherosclerosis is in high expression.

The present invention also provides the existing kit of detection TM4SF19 protein or diagnostic device in preparation for diagnosing And/or the application in the medicament of disease caused by prevention and/or treatment atherosclerosis, the kit or diagnostic device are used The in-vitro diagnosis and/or risk stratification of the disease caused by atherosclerosis.

Optionally, the reagent is used to measure the expression quantity of TM4SF19 protein in humoral sample.

Optionally, the reagent is exempted from by immunoblotting, enzyme linked immunosorbent assay (ELISA), radiommunoassay, radiation Epidemic disease diffusion method, Auchterlonie immunodiffusion, rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation analysis, complement knot One or more of analytic approach, fluidic cell fluorescence point side technology and protein-chip method is closed to detect TM4SF19 protein Expression quantity.

Optionally, disease caused by the expression quantity of TM4SF19 protein and atherosclerosis is tight in the humoral sample Weight degree is positively correlated.

Optionally, the humoral sample is selected from serum.

Optionally, the medicament be used for diagnose or monitor disease caused by atherosclerosis presence and/or process and/ Or severity and/or prognosis.

The present invention also provides the DNA or its mRNA for Expression of TM 4SF19 protein or the homologue for possessing its function to make For application of the biomarker in the diagnostic reagent for preparing or screening disease caused by atherosclerosis.

The present invention also provides a kind of composition for diagnosing atherosclerotic, comprising with TM4SF19 protein or its Fragments specific in conjunction with antibody, antigen-binding fragment or more skins or with core that the TM4SF19 protein is encoded Probe, primer sets or the nucleotide of nucleotide sequence specific binding.

The present invention also provides a kind of kits, the reagent comprising above-mentioned composition and for detecting the composition.

As described above, it is of the invention for the molecular marker TM4SF19 of diagnosing atherosclerotic and its application, have Below the utility model has the advantages that present invention discover that can predict that artery is athero- by the TM4SF19 protein concentration in detection blood samples of patients The generation of disease caused by hardening, or above-mentioned disease is diagnosed, risk stratification etc..

Detailed description of the invention

Fig. 1 .1 is shown as the genetic chip thermal map of atherosclerotic tissue and normal person's comparison.

Fig. 1 .2 is shown as the gene chip results figure of atherosclerotic tissue and normal person's comparison.

Fig. 2 .1 is shown as smooth muscle cell TM4SF19 in oxldl stimulation and changes over time the increased signal of expression quantity Figure.

Fig. 2 .2 be shown as smooth muscle cell oxldl stimulation when TM4SF19 with concentration change the increased signal of expression quantity Figure.

Fig. 2 .3 is shown as macrophage TM4SF19 in oxldl stimulation and changes over time the increased schematic diagram of expression quantity.

Fig. 2 .4 be shown as macrophage oxldl stimulation when TM4SF19 with concentration change the increased schematic diagram of expression quantity.

Fig. 2 .5 is shown as endothelial cell TM4SF19 in LPS stimulation and changes over time the increased schematic diagram of expression quantity.

Fig. 2 .6 be shown as endothelial cell LPS stimulation when TM4SF19 with concentration change the increased schematic diagram of expression quantity.

Fig. 3 is shown as endothelial cell TM4SF19mRNA in LPS stimulation and changes over time the increased schematic diagram of expression quantity.

Fig. 4 is shown as the result schematic diagram of ELISA detection TM4SF19 serum specimen concentration.

Specific embodiment

Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification Other advantages and efficacy of the present invention can be easily understood for disclosed content.The present invention can also pass through in addition different specific realities The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints and application, without departing from Various modifications or alterations are carried out under spirit of the invention.

Cross-film -4-L6 family protein 19 (Transmembrane 4L Six Family Member 19, TM4SF19), again Name OCTM4, belongs to a member in cross-film -4-L6 family protein (TM4SFs), molecular weight 22kD is encoded by TM4SF19 gene. TM4SF19 is positioned at 3q29, contains 3 exons, the mRNA sequence of total length 14838bp, TM4SF19 such as SEQ ID NO.1 altogether Shown, amino acid sequence is as shown in SEQ ID NO.2.TM4SF19 is higher in testis and esophagus expression, should in human brain tissue Gene high level expression in top, occipital lobe, hippocampus and cerebellum.Family protein member passes through the interaction with integrin It works in various cell processes, including cell Proliferation, movement and adherency.

TM4SF19 albumen not yet someone reports, and specific effect waits to define, and early-stage study is shown, TM4SF19 may pass through Inflammation and cholesterol metabolic disorder and the occurrence and development for promoting atherosclerosis.

TM4SF19 forward primer sequence is as shown in SEQ ID NO.3.

TM4SF19 forward primer sequence is as shown in SEQ ID NO.4.

1, experimental method

(1) cell culture

(the DMEM culture of 4ml complete medium is added in Tissue Culture Dish in recovery Human umbilical vein endothelial cells (HUVEC) + 10% fetal calf serum of base), be placed in cell incubator (37 DEG C, 5%CO₂) it is incubated for culture.After cell confluency degree up to after 80~90% Cell is taken out, 500 μ l trypsin digestion cells are added, when microscopically observation cell starts shrinkage, 200 μ l fetal calf serums are added Digestion is terminated, cell is gently blown and beaten repeatedly, it is made to fall off from ware bottom, goes to 15ml centrifuge tube, 1300r/min is centrifuged 5min, abandons Supernatant is removed, appropriate complete medium is added, cell is resuspended, equivalent is dispensed into 2-4 new Tissue Culture Dish, with complete culture Base supplies 4ml/ ware, continues at and is incubated for culture in cell incubator.Above step is repeated, cell is kept normally to pass on, in case after Continuous experiment uses.

(2) intracellular total protein extraction

Cell to be processed is taken out from cell incubator, discards culture solution, is washed 2 times with PBS, PBS is discarded, is added 80 μ l protein lysate makes lysate cover ware bottom, stands 5min and scrapes cell with cell scraper after cell is sufficiently cracked, uses Sample loading gun is drawn whole cell suspensions and is transferred in 1.5ml EP pipe, and 4 DEG C, 13000r/min centrifugation 3min collect supernatant simultaneously It is transferred in the new 1.5ml EP pipe marked, is saved after quantitative in -20 DEG C stand-by.

(3) intracellular Total RNAs extraction

Cell to be processed is taken out from cell incubator, discards culture solution, is washed 2 times with PBS, discards PBS, is added 500 μ l RNAiso Plus lytic cells can be completely covered entire ware bottom, be drawn and be cracked with sample loading gun after 5-10min Liquid blows and beats cell repeatedly, so that it is fallen off from ware bottom, stands 5min on ice, and cell pyrolysis liquid is all collected and transferred to In 1.5mlEP pipe and mark.It is added chloroform (the 1/5 of RNAiso Plus volume), covers tightly EP pipe lid, quickly acutely It turns upside down and vibrates 15s, come into full contact with organic phase with water phase, be stored at room temperature 2-3min, until after emulsifying soln is abundant, room temperature 5min is stood, 4 DEG C, 12000g-14000g centrifugation 15min take out EP pipe, it is seen that be divided into three layers: upper layer is colourless aqueous phase, is rich in Cell RNA；Middle layer is white layer, wherein containing a large amount of protein；Bottom is red organic phase.Slowly gently with sample loading gun A layer supernatant is sucted, is transferred in the new 1.5ml EP pipe marked.The isopropanol isometric with the supernatant of absorption is added, gently After soft mixing of turning upside down, it is stored at room temperature 10min, 4 DEG C, 12000g centrifugation 10min abandon supernatant, 75% ethanol washing are added RNA, has flicked EP tube bottom precipitating, and 4 DEG C, 12000g centrifugation 5min abandon supernatant, repeated washing is primary, dries RNA precipitate at room temperature After becoming clear, colorless substance to white precipitate, 20 μ l DEPC water are added in 10-15min, and quantitative latter -80 DEG C save for use.

(4) real-time fluorescence quantitative PCR (qPCR)

1. using Takara Real Time RNA Reverse Transcriptase kit by the total serum IgE reverse transcription of extraction be cDNA, system It is as shown in the table:

Table 1

Reverse transcription reaction condition are as follows: 37 DEG C, 15min → 85 DEG C, 5s → 4 DEG C maintain, and the mRNA obtained through reverse transcription is anti- Liquid is answered to dilute 5 times, to react for subsequent PCR.

2. carrying out PCR reaction using Takara real-time fluorescence quantitative PCR kit, system is as shown in the table:

Table 2

3. being carried out amplification reaction using Cobas Z480 real-time PCR, condition after the completion of system is prepared are as follows: 90 DEG C, 10min → [95 DEG C, 10s → 60 DEG C, 60s is recycled 40 times] → 95 DEG C, 1min → 60 DEG C, 30s → 40 DEG C, 30s.

Real-time fluorescence quantitative PCR reaction is internal reference with GAPDH, and uses 2^-ΔΔCtCalculate the opposite table of target gene mRNA Up to amount.

(5) protein immunoblotting test (Western Blot)

1. preparative separation glue and concentration glue, system are as shown in the table.

10% separation gel system is as shown in table 3:

Table 3

5% concentrating colloidal system is as shown in table 4:

Table 4

2. installing the gel prepared into Vertial electrophorestic tank, 5 μ l marker instruction is added in the first hole swimming lane Agent, remaining each hole swimming lane are separately added into 80 μ g protein samples (volume is depending on each sample concentration), power on, 80V electrophoresis 20min is in spacer gel and separation gel line of demarcation to protein sample, voltage is then adjusted to 120V, electrophoresis 1h, albumen is in voltage It gradually shifts, and successively separates to different level position down under effect.According to marker and protein molecular size, to required egg It is white reach can fully disengaged position when, stop electrophoresis.

3. taking out the gel in electrophoresis tank, the corresponding band of destination protein is cut according to marker, opens transferring film folder, is being turned Film folder black side successively puts filter paper, gel-tape, pvdf membrane, and filter paper is gently pressed on pvdf membrane back and forth with passivity plate level It presses to drive the bubble of the inside away, then covers transferring film plate white board, the black flour of transferring film plate is directed at the black flour of transferring film slot, transferring film plate Fine flour is against the red face of slot.Transferring film slot is filled into transferring film liquid, is immersed in gel where all purposes albumen in transferring film liquid, then Transferring film slot is placed in mixture of ice and water, power supply is opened, 100V, 70min carry out transferring film.

4. taking out the band after transferring film, it is placed in 5% skim milk, closes 2h in shaking table room temperature.

5. taking out band, cleaned 3 times with TBST, each 5min.

6. prepare with the corresponding antibody of destination protein, band is put into antibody in solution, was incubated at a slow speed in 4 DEG C of shaking tables Night.

7. taking out band, cleaned 3 times with TBST, each 5min.

8. preparing corresponding secondary antibody (dilution ratio 1:5000-1:10000) according to primary antibody kind, band is moved into secondary antibody In solution, 1h is slowly incubated on room temperature shaker.

9. taking out band, cleaned 3 times with TBST, each 5min.

10. being taken in item and super quick luminescent solution being added dropwise, it is exposed in exposure instrument and analyzes result.

2, analysis of experimental results

2.1 genetic chip

Genetic chip (Fig. 1 .1, Fig. 1 .2) display, the expression of patient group TM4SF19 is more right in artery plaque tissue According to group 223 times of a up-regulation.

2.2Western blot

1. with the OxLDL ELISA (Ox-ldl) of 50 μ g/ml concentration stimulate respectively smooth muscle cell (SMC) 0h, 12h, for 24 hours and 48h, measures expression of the TM4SF19 in cell, experimental result is as shown in Fig. 2 .1, the results showed that The expression of TM4SF19 rises in smooth muscle cell with the raising of ox-LDL stimulation time.

2. being stimulated respectively with the OxLDL ELISA (Ox-ldl) of 0,25 μ g/ml, 50 μ g/ml and 100 μ g/ml concentration Smooth muscle cell (SMC) 48h measures expression of the TM4SF19 in cell, and experimental result is as shown in Fig. 2 .2, the results showed that The expression of TM4SF19 rises in smooth muscle cell with the raising of ox-LDL irritaiting concentration.

3. stimulating monocytes/macrophages (Thp-1) respectively with the OxLDL ELISA (Ox-ldl) of 50 μ g/ml concentration 0h, 12h, for 24 hours and 48h, measures expression of the TM4SF19 in cell, experimental result is as shown in Fig. 2 .3, the results showed that The expression of TM4SF19 rises in macrophage with the raising of ox-LDL stimulation time.

4. being stimulated respectively with the OxLDL ELISA (Ox-ldl) of 0,25 μ g/ml, 50 μ g/ml and 100 μ g/ml concentration Smooth muscle cell (SMC) 48h measures expression of the TM4SF19 in cell, and experimental result is as shown in Fig. 2 .4, the results showed that The expression of TM4SF19 rises in macrophage with the raising of ox-LDL irritaiting concentration.

5. with the lipopolysaccharides (LPS) of 500ng/ml concentration respectively stimulating endothelial cell (EC) 0h, 12h, for 24 hours, 36h and 48h, Expression of the TM4SF19 in cell is measured, experimental result is as shown in Fig. 2 .5, the results showed that the expression of TM4SF19 is in endothelium Rise in cell with the raising of LPS stimulation time.

6. being stimulated respectively with the lipopolysaccharides (LPS) of 0,100ng/ml, 250ng/ml, 500ng/ml and 1000ng/ml concentration Endothelial cell (EC) 48h measures expression of the TM4SF19 in cell, and experimental result is as shown in Fig. 2 .6, the results showed that The expression of TM4SF19 rises in endothelial cell with the raising of LPS irritaiting concentration.

3、PCR

With the lipopolysaccharides (LPS) of 500ng/ml concentration respectively stimulating endothelial cell (EC) 0h, 1.5h, 3h, 6h, 12h, for 24 hours, 36h and 48h, measures expression of the TM4SF19 in cell, and experimental result is as shown in Figure 3, the results showed that the table of TM4SF19 Rise in early days with the raising of LPS stimulation time up in endothelial cell, peak value was reached at 12 hours, subsequent expression restores Normally.

4、ELISA

[normal control (NC group), simple diabetes group (DM group), moves simple atherosclerosis group (AS group) for four groupings Pulse atherosclerosis complicated with diabetes group (AS+DM group)] expression (pg/ml) of TM4SF19 in serum sample, experimental result is such as Shown in Fig. 4, as a result, it has been found that, NC group is compared, the expression quantity of the TM4SF19 in AS+DM group significantly rises, there is statistical difference, And the expression of TM4SF19 and NC group no significant difference in DM group, illustrate TM4SF19 high expression in atherosclerotic, And do not interfered by other chronic diseases, i.e. TM4SF19 is the marker of a potential diagnosing atherosclerotic.

In conclusion present invention discover that can be predicted by the TM4SF19 protein concentration in detection patient serum sample The generation of disease caused by atherosclerosis, or above-mentioned disease is diagnosed, risk stratification etc..

The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as At all equivalent modifications or change, should be covered by the claims of the present invention.

SEQUENCE LISTING

<110>Hospital of Southern Medical University

<120>for the molecular marker TM4SF19 of diagnosing atherosclerotic and application

<130> PCQNF192605

<160> 4

<170> PatentIn version 3.5

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<211> 1073

<212> DNA

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<223>mRNA of TM4SF19

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acgtatatac agagcctccc tggccctcct ggaaagagtc ctggaaagac aaccttcagg 60

tccagccctg gagctggagg agtggagccc cactctgaag acgcagcctt tctccaggtt 120

ctgtctctcc cattctgatt cttgacacca gatgcaggat ggtgtcctct ccctgcacgc 180

aggcaagctc acggacttgc tcccgtatcc tgggactgag ccttgggact gcagccctgt 240

ttgctgctgg ggccaacgtg gcactcctcc ttcctaactg ggatgtcacc tacctgttga 300

ggggcctcct tggcaggcat gccatgctgg gaactgggct ctggggagga ggcctcatgg 360

tactcactgc agctatcctc atctccttga tgggctggag atacggctgc ttcagtaaga 420

gtgggctctg tcgaagcgtg cttactgctc tgttgtcagg tggcctggct ttacttggag 480

ccctgatttg ctttgtcact tctggagttg ctctgaaaga tggtcctttt tgcatgtttg 540

atgtttcatc cttcaatcag acacaagctt ggaaatatgg ttacccattc aaagacctgc 600

atagaattat ctgtatgacc gttcgctctg gaactccgtc tgcctggagc cctctgcagc 660

tgttgtctgg cacgtgtccc tcttctccgc ccttctgtgc atcagcctgc tccagcttct 720

cctggtggtc gttcatgtca tcaacagcct cctgggcctt ttctgcagcc tctgcgagaa 780

gtgacaggca gaaccttcac ttgcaagcat gggtgttttc atcatcggct gtcttgaatc 840

ctttctacaa ggagtgggta cgaattataa acaaacttcc cctttaggta tccctggagt 900

aataatgaca acaaaattca ctgcaggtcg gtggaatgat agaatgcatt ttaaatcaca 960

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<223>amino acid sequence of TM4SF19

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Met Val Ser Ser Pro Cys Thr Gln Ala Ser Ser Arg Thr Cys Ser Arg

1 5 10 15

Ile Leu Gly Leu Ser Leu Gly Thr Ala Ala Leu Phe Ala Ala Gly Ala

20 25 30

Asn Val Ala Leu Leu Leu Pro Asn Trp Asp Val Thr Tyr Leu Leu Arg

35 40 45

Gly Leu Leu Gly Arg His Ala Met Leu Gly Thr Gly Leu Trp Gly Gly

50 55 60

Gly Leu Met Val Leu Thr Ala Ala Ile Leu Ile Ser Leu Met Gly Trp

65 70 75 80

Arg Tyr Gly Cys Phe Ser Lys Ser Gly Leu Cys Arg Ser Val Leu Thr

85 90 95

Ala Leu Leu Ser Gly Gly Leu Ala Leu Leu Gly Ala Leu Ile Cys Phe

100 105 110

Val Thr Ser Gly Val Ala Leu Lys Asp Gly Pro Phe Cys Met Phe Asp

115 120 125

Val Ser Ser Phe Asn Gln Thr Gln Ala Trp Lys Tyr Gly Tyr Pro Phe

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Lys Asp Leu His Arg Ile Ile Cys Met Thr Val Arg Ser Gly Thr Pro

145 150 155 160

Ser Ala Trp Ser Pro Leu Gln Leu Leu Ser Gly Thr Cys Pro Ser Ser

165 170 175

Pro Pro Phe Cys Ala Ser Ala Cys Ser Ser Phe Ser Trp Trp Ser Phe

180 185 190

Met Ser Ser Thr Ala Ser Trp Ala Phe Ser Ala Ala Ser Ala Arg Ser

195 200 205

Asp Arg Gln Asn Leu His Leu Gln Ala Trp Val Phe Ser Ser Ser Ala

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Val Leu Asn Pro Phe Tyr Lys Glu Trp Val Arg Ile Ile Asn Lys Leu

225 230 235 240

Pro Leu

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<223>TM4SF19 forward primer

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cacggacttg ctcccgtatc 20

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gcccctcaac aggtaggtg 19

Claims (10)

1. cross-film -4-L6 family protein 19 or its peptide fragment are being prepared or are being screened caused by atherosclerosis as biomarker Application in reagent for disease diagnosis, the diagnostic reagent are used for the in-vitro diagnosis and/or danger of disease caused by atherosclerosis Danger layering.

2. application according to claim 1, it is characterised in that: disease caused by the pulse atherosclerosis includes cardiovascular disease Disease；And/or the TM4SF19 protein level of Disease caused by atherosclerosis is in high expression.

3. the existing kit or diagnostic device that detect TM4SF19 protein are being prepared for diagnosing and/or preventing and/or control The application in the medicament of disease caused by atherosclerosis is treated, the kit or diagnostic device draw for atherosclerosis The in-vitro diagnosis and/or risk stratification of the disease risen.

4. application according to claim 3, it is characterised in that: the reagent is for measuring TM4SF19 egg in humoral sample The expression quantity of white matter.

5. application according to claim 4, it is characterised in that: the expression quantity of TM4SF19 protein in the humoral sample It is positively correlated with disease severity caused by atherosclerosis；

And/or the humoral sample is selected from serum.

6. application according to claim 3, it is characterised in that: the reagent is inhaled by immunoblotting, enzyme linked immunological Attached measurement, radiommunoassay, radioimmunodiffusion, Auchterlonie immunodiffusion, rocket immunoelectrophoresis, immunohistochemistry Dyeing, immunoprecipitation analysis, complement fixation assays method, fluidic cell fluorescence divide one of side technology and protein-chip with Upper method detects the expression quantity of TM4SF19 protein.

7. application according to claim 3, it is characterised in that: the medicament draws for diagnosing or monitoring atherosclerosis The presence of the disease risen and/or process and/or severity and/or prognosis.

8. the DNA or its mRNA that are used for Expression of TM 4SF19 protein or the homologue for possessing its function exist as biomarker Application in the diagnostic reagent of disease caused by preparation or screening atherosclerosis.

9. the composition of diagnosing atherosclerotic is used for, comprising anti-in conjunction with TM4SF19 protein or its fragments specific It body, antigen-binding fragment or more skins or is specifically bound with the nucleotide sequence that is encoded to the TM4SF19 protein Probe, primer sets or nucleotide.

10. a kind of kit for diagnosing atherosclerotic, comprising composition as claimed in claim 9 and for detecting State the reagent of composition.