CN107164546A - MiRNA, composition and its application in diagnosing the illness - Google Patents

Abstract

Application the present invention relates to miRNA, composition and its in diagnosing the illness, is specifically related to the application of 5p, the miR 1262 of miR 942 and their composition in diagnosing myocardial infarction.MiRNA take part in during the different pathological of cardiovascular reconstruction, including myocardial hypertrophy, fibrosis, myocardial function change and angiogenesis etc., miRNA dynamic change is detected, may be given a clue for generation, the development of disease, and then instruct clinical early intervention, the effectively development of control disease.The 5p of miR 942 and miR 1262 and myocardial infarction that the present invention is studied are closely related, can individually or merge for myocardial infarction clinical diagnosis and prevention detection, be clinically dependent diagnostic reagent research and development lay the foundation.

Description

MiRNA, composition and its application in diagnosing the illness

Technical field

The present invention relates to medical diagnosis on disease field, and in particular to miRNA, composition and its application in diagnosing the illness, more It is specifically related to the application of miR-942-5p, miR-1262 and their composition in diagnosing myocardial infarction.

Background technology

Recent studies suggest that, miRNA participates in the transcript and expression regulation and control of 30%-50% genes, in the life of cardiovascular system Very important effect is all played in reason and pathomechanism.Existing report shows that miRNA take part in the difference disease of cardiovascular reconstruction During reason, including the change of myocardial hypertrophy, fibrosis, myocardial function and angiogenesis etc., because miRNA change is earlier than base The change of cause and albumen, also earlier than the appearance of disease symptomses, thus detects miRNA dynamic change, it is possible to be the hair of disease Raw, development is given a clue, and then instructs clinical early intervention, the effectively development of control disease.Acute myocardial infarction AMI (AMI) is most tight Weight a kind of angiocardiopathy, early stage, accurately diagnosis can ensure reperfusion as treatment immediately begin to be likely to reduced the death rate. The application of some biomarkers such as cardiac muscle troponin I and diagnosis AMI, can improve current diagnosis, be acute myocardial infarction AMI New method.Conventional research shows that miR-30a is related to myocardial hypertrophy, can be used as diagnosing acute myocardial infarction biomarker Potential clinical value, but these are also far from enough, and clinical accurate diagnosis and treatment needs more candidate molecules marks.

The present invention is based on high-flux sequence method, obtains myocardial infarction miRNA expression data, meanwhile, with reference to GEO data Storehouse myocardial infarction correlation miRNA data, carry out bioinformatic analysis checking, miR- are picked out from the miRNA of candidate 942-5p and miR-1262 carry out molecular biology checking, as a result show, miR-942-5p and miR-1262 and myocardial infarction are close Cut is closed.Can individually or merge for myocardial infarction clinical diagnosis and prevention detection, be clinically dependent diagnostic reagent research and development Lay the foundation.

The content of the invention

Detect application of the miRNA preparation in diagnosing myocardial infarction reagent is prepared, the miRNA in following one Plant or several：miR-942-5p、miR-1262.

The sequence of the miR-942-5p is shown in sequence table SEQ ID NO 1.

The sequence of the miR-1262 is shown in sequence table SEQ ID NO 2.

It is preferred that, joint-detection miR-942-5p and miR-1262 preparation answering in diagnosing myocardial infarction reagent is prepared With.

Further, diagnosing myocardial infarction reagent include based on high-flux sequence method and/or based on quantifying PCR method and/ Or based on miR-942-5p and/or miR-1262 in probing procedure detection sample.

It is preferred to use northern hybridizing methods, miRNA chip of expression spectrum, ribozyme protection analytical technology, RAKE methods, original position MiR-942-5p and/or miR-1262 transcription in hybridization, bead-based flow-cytometry detection sample.

Described sample is peripheral blood.

Further, Risk of myocardial infarction increase is suffered from the expression display of miR-942-5p and/or miR-1262 height in sample.

It is preferred that, specific amplification miR-942-5p and/or miR-1262 primer are included based on quantifying PCR method, is entered Preferably, specific amplification miR-942-5p primer sequence is SEQ ID NO 3, specific amplification miR-1262 primer to one step Sequence is SEQ ID NO 4；Nucleic acid array hybridizing with miR-942-5p and/or miR-1262 is included based on probing procedure Probe.

Further, the miRNA is miR-942-5p and/or miR-1262 precursor, and precursor sequence is SEQ ID NO 5 With SEQ ID NO 6.

It is an object of the invention to provide the transcription for lowering miR-942-5p and/or miR-1262 and/or suppression miR- 942-5p and/or miR-1262 active reagent is preparing the application in preventing and treating myocardial infarction preparation.

It is preferred that, using ASON, antagomiRs, miRNA sponge, miRNA Erasers, Target The method of Masking and/or Mutiple Targets ASON lowers miR-942-5p and/or miR-1262 transcription and/or resistance Disconnected miR-942-5p and/or miR-1262 activity.

Diagnosis is being prepared it is an object of the invention to provide detection miR-942-5p and/or miR-1262 target gene preparation Application in myocardial infarction preparation.

The target gene be DIANA-MICROT, MICRORNA.ORG, MIRDB, RNA22-HAS, TARGETMINER, TARGETSCAN-VERT databases show miR-942-5p and/or miR-1262 target gene.

Definition：

The method of detection miRNA expression is mainly included based on high throughput sequencing technologies, based on nucleotides at this stage Hybridization and the miRNA detection methods of PCR-based.MiRNA detection methods based on probe hybridization technique are a kind of direct Detection Methods, Sample rna need not in advance be expanded, including northern hybridizing methods, miRNA chip of expression spectrum, ribozyme protection analysis skill The technologies such as art, RAKE methods, in situ hybridization, bead-based flow-cytometry.

(1) Northern hybridizes

Also known as Northern blot is most classical detection eucaryote RNA sizes, estimates the experimental method of its abundance.Base Present principles are as follows：MiRNA samples are fixed on carrier (such as silicon chip, microballoon or film) first, then it is miscellaneous with the probe of process mark Hand over, signal detection is carried out after washing unnecessary hybridization probe；Can also the first fixed and target miRNA sequence complementation on carrier DNA probe, then with the sample miRNA hybridization by mark, then carries out signal detection.The method of signal mark includes isotope Mark, fluorescence labeling and nano gold mark etc..

(2) miRNA chip of expression spectrum

Principle is equally that the target molecule on solid support is detected using label probe.Pass through miRNA in design chips Gene and internalcontrol sequence, can Accurate Analysis go out the expression of corresponding miRNA in sample.Genetic chip has high-throughout excellent Point, can once detect whole expression of hundreds of genes in same sample.The liquid-phase chip that Luminex companies develop (Liquid chip) is also known as multifunctional suspending dot matrix (Multi analyte suspension array, MASA), is new Generation biochip technology.Liquid-phase chip system is made up of many spherulas for main matrix, is fixed with not on every kind of spherula With probe molecule, in order to distinguish different probes, sphere matrix that each is used for label probe is all unique with one Color numbers, these spherulas are suspended in a liquid-phase system, liquid-phase chip system is just constituted.The system can be to same Multiple different moleculars in one trace sample carry out quick qualitative and quantitative analysis simultaneously, and this detection technique is referred to as FMAP (Flexible multianalyte profiling) technology.Molecule hybridization is carried out in aaerosol solution, detection speed pole It hurry up.

(3) ribozyme protection analytical technology (RPA)

MiRNA detection can also protect analytical technology using ribozyme, and the probe marked and RNA samples to be measured are mixed Close, hybridize after thermal denaturation, non-hybridized RNA and the single-chain nucleic acid enzymic digestion of unnecessary probe, purify after heat inactivation nuclease by The RNA molecule of protection, finally by denaturation PAGE electrophoretic separation probes, colour developing.This new method based on solution hybridization is simple Quickly, sensitivity is high, but is also only used for analyzing known miRNA.

(4) RAKE methods

RAKE methods (RNA primed array based Klenow emzyme) are the bases in miRNA microarray The Klenow fragments of DNA polymerase i are utilized on plinth, make miRNA and the method for fixed DNA probe hybridization.RAKE can be sensitive Specifically detect miRNA, it is adaptable to largely quickly screen all miRNA that oneself knows.Can be in specific cell and tumour Detect miRNA express spectra situations.Moreover, RAKE methods can also be from the tissue of the FFPE secured by formalin Isolate miRNA and it is analyzed, be the door that analysis miRNA opens hope from sample is achieved.

(5) in situ hybridization (in situ hybridization)

Hybridization in situ technique can intuitively understand miRNA expression ways, be a kind of easier of observation miRNA spatial and temporal expressions Method, normal mark mode includes digoxin, biotin, fluorescence labeling etc..In situ hybridization (Locked on the basis of locked nucleic acid Nucleic Acid (LNA) based in situ hybridization (LNA-ISH)) it is the more probe side of current application Formula.

(6) bead-based flow-cytometry (bead-based flow cytometry)

It is a kind of liquid-phase chip technology, FCM analysis is organically combined, had concurrently by this method with chip technology Flux is big, detection speed is fast, sensitivity is high and it is specific good the features such as.

(7) Real-Time Fluorescent Quantitative PCR Technique (Real-time PCR, RT-PCR)

Fluoroscopic examination PCR instrument can draw dynamic changing curve to the cumulative speed of extension increasing sequence during whole PCR.Anti- Answer the initial concentration of target sequence in mixed system bigger, it is desirable to which the PCR cycle number for obtaining amplified production specific output is (general to use Specific threshold period Ct is expressed) it is fewer.Because miRNA length is only 22nt, it is such that traditional qRT-PCR is not suitable for amplification Short fragment.There are several real time quantitative PCR methods for miRNA, such as tailing method, neck ring method now.Neck ring method is a kind of Preferable miRNA detections qRT-PCR methods：Special loop-stem structure primer is designed first, using miRNA to be measured as template reverse transcription The chains of cDNA first are synthesized, the cDNA one end is stem Loop primer, and stem loop structure, which is opened, substantially increases cDNA length, then Real-time quantitative PCR amplification is carried out by template design primer of the cDNA of synthesis.QRT-PCR has specificity height, sensitivity good, fast A variety of advantages such as fast simple.

(8) PCR sequencing PCR

Most of known miRNA is to be sequenced to find and identify by cDNA clone.The method needs first to build miRNA CDNA library, then enter performing PCR amplification, amplified production be then cloned on expression vector be sequenced.Takada develops one kind and changed The amplification PCR cloning PCR (miRNA amplification profiling, mRAP) entered, mRAP methods are first connected at miRNA 3 ' ends Joint, then with the reverse transcription primer reverse transcription complementary with joint.Because specific reverse transcriptase has end deoxynucleotide Transferase active, some nucleotides (mainly deoxycytidylic acid) can be connected to 3 ' ends of the cDNA chains that reverse transcription goes out.When 5 ' End connector is the achievable PCR expansions to cDNA with after poly (C) cohesive end annealing of cDNA chains, adding a pair of general primers Increase.Due to mRAP High sensitivities, the expression quantity of miRNA in a small amount of tissue can be directly detected with clone and sequencing technologies.Label Sequence PCR cloning PCR is a kind of to have developed detection efficiency on the basis of serial analysis of gene expression (SAGE) technology higher MiRAGE (miRNA SAGE) PCR cloning PCR, the method sub-series big by generating can detect multiple by single sequencing reaction MiRNA, hence it is evident that improve detection efficiency.

High-flux sequence (High-throughput sequencing) is also known as sequencing technologies (next of future generation Generation sequencing) it is the change to tradition sequencing revolution, once to hundreds of thousands to millions of DNA Molecule carries out sequencing, greatly improves sequencing efficiency.This kind of large scale sequencing technology greatly improves multiple species and lost The solution reading rate of information is passed, to obtain all miRNA sequence information, decryption miRNA collection of illustrative plates, which is provided, to be ensured.While high flux The analysis for make it that careful overall picture is carried out to the transcript profile and genome of a species is sequenced, so the depth that is otherwise known as It is sequenced (deep sequencing).The representative of high-flux sequence platform is 454 sequenator (Roch of Roche Holding Ag (Roche) GSFLX sequencer), the Solexa genome analysises instrument (Illumina Genome Analyzer) of Illumina companies and ABI SOLiD sequenators (ABI SOLiD sequencer).

MicroRNA gain-of-functions technology based on RNA be by exogenous supplement miRNAs synthesis precursor substance come Raise miRNAs level.For example, can be with the artificial synthesized bob folder sample RNA (short consistent with endogenous miRNA sequence Hairpin RNA, shRNA), promoter is done by polymerase II or III, with virus for carrier transfectional cell, by Dicer enzyme modifications RISC is loaded into afterwards to play a role, it is lasting equivalent to rise pre-miRNA level, action effect stabilization.

This technique avoids the nonspecific action of miRNA and gene for gene specific miR Mimics technologies.This people The specific oligonucleotide chain combined with the UTR of target gene 3 ' complementations of work synthesis, can be played with being adjusted after the transcription of miRNA identicals Section is acted on.

The major way of miRNA regulation and control has two kinds：A kind of is the not fully complementary pairings of 3 ' UTR with target gene mRNA, resistance The translation of disconnected target gene, so that regulatory gene is expressed；It is another be it is similar with siRNA, when miRNA and mRNA complete complementaries are matched When, Ago2 albumen directly results in its degraded by cutting mRNA, realizes gene silencing.In a word, miRNA is presently believed to which kind of side Formula is acted on target gene and miRNA is relevant with the pairing degree of target gene.When miRNA matches incomplete with target gene, MiRNA is just played a role with the expression for suppressing target gene；When miRNA matches complete with target gene section sequence, it is possible to Target gene is caused to be broken in complementary region and cause gene silencing.

Brief description of the drawings

Fig. 1 is the ROC curve figure of miR-942-5p diagnosing myocardial infarctions

Fig. 2 is the ROC curve figure of miR-1262 diagnosing myocardial infarctions

Fig. 3 is the ROC curve figure of miRNA Combining diagnosis myocardial infarctions

Fig. 4 is miR-942-5p and miR-1262 relative expression's spirograms

Embodiment

With reference to specific embodiment, the present invention is expanded on further, is only used for explaining the present invention, and it is not intended that to this The limitation of invention.It will be understood by those skilled in the art that：Can in the case where not departing from the principle and objective of the present invention So that these embodiments are carried out with a variety of change, modification, replacement and modification, the scope of the present invention is limited by claim and its equivalent It is fixed.The experimental method of unreceipted actual conditions in the following example, generally according to normal condition or according to the bar proposed by manufacturer Part examinations.

The collection of the sample of embodiment 1 and Total RNAs extraction

Hospital in March, 2015 is collected each 6 to the myocardial infarction patient of in September, 2016 peripheral bloods and normal healthy controls peripheral blood. Diagnosis of Acute Myocardial Infarction standard：According to third time in 2012《Myocardial infarction whole world unified definition》The diagnostic criteria system of recommendation It is fixed.Detect myocardial necrosis marker (mainly troponin) level to rise and/or decline, at least 1 time more than reference value 99%th percentile of the upper limit, and at least with the symptom of a following ischemic.

1) symptom of myocardial ischemia；

2) the ischemic ECG of kainogenesis changes；

3) there are pathologic Q ripples in electrocardiogram；

4) radiological evidence shows new myocardial activity and lost or new regional wall motion abnormality；

5) coronarography or postmortem find there is fresh thrombus in coronary artery.

Blood rna extraction standard：RNA purity：OD260/280≤1.8,28S/18S≤1；RNA integralities：RIN Zhi≤ 7.0.RNA integrality detection methods：(agarose coagulates for Agilent 2100 (the Nano kit of RNA 6000), agarose gel electrophoresis Gum concentration：1% agarose gel；Voltage：5V/cm；Time：20min).

Embodiment 2 is sequenced, data analysis and electronically validating

Sequencing：MiRNA is sequenced with llumina Hiseq2500/Miseq second generation high throughput sequencing technologies, By go joint, go low quality, the process such as depollute completes the processing of data, obtains final data.Pass through transcript profile data point Analysis software carries out progress t-test after background correction to miRNA sequencing initial data and obtains P values, is then examined and closed using Fisher And P values, screening differential expression miRNA.

It is final to be selected from the differential expression miRNA of candidate to miR-942-5p, miR-1262, miR-520c-3p, carry out Later experiments are verified.

The electronically validating of embodiment 3

Electronically validating：3 sets of mRNA data sets are obtained from the screening of GEO (Gene Expression Omnibus) database (GSE48060, GSE34198 and GSE61145-GPL6884) and 2 sets of miRNA data sets (GSE61741, GSE31568), 3 sets MRNA data sets (GSE48060, GSE34198 and GSE61145-GPL6884) have gene 13680, we by using MetaMA bags, which are calculated and merged using merging P value methods, obtains 612 (FDR after effect value<0.05) difference expression gene, wherein on 352 are adjusted, 260 are lowered；The common miRNA numbers 848 of 2 sets of data collection (GSE24371, GSE17498).Use R bags MetaMA merges the analysis of P values to two sets of miRNA data sets, finds 15 differential expression miRNA (FDR<0.001&&| Combined.ES|>1).As a result show that electronically validating result is consistent with sequencing result expression trend.

It is to set up Receiver Operating Characteristics for single miRNA molecule or the method for the efficiency evaluation of diagnostic model (receiver operating characteristic, ROC) curve, passes through (the Area Under of area under calculated curve Curve) come judge diagnosis ability.Area value under ROC curve is between 1.0 and 0.5, in AUC>In the case of 0.5, AUC Closer to 1, illustrate that diagnosis effect is better, AUC has relatively low accuracy in 0.5-0.7, AUC is fixed in 0.7-0.9 True property, has high accuracy when AUC is more than 0.9.We will utilize GSE31568 data sets (including 70 control groups and 20 Example case group), and then analyzed, as a result show, the AUC of miR-942-5p diagnosing myocardial infarctions is 0.783 (see Fig. 1), The AUC of miR-1262 diagnosing myocardial infarctions is 0.748 (see Fig. 2), and the AUC of miR-520c-3p diagnosing myocardial infarctions is 0.729, Show that its accuracy rate is higher, show three can as diagnosing myocardial infarction molecular marker, the analysis of further Combining diagnosis (see Fig. 3), the AUC of miR-520c-3p and miR-1262 Combining diagnosis myocardial infarctions is 0.776, miR-1262 and miR-942- The AUC of 5p Combining diagnosis myocardial infarctions is 0.805, miR-520c-3p, miR-1262 and miR-942-5p Combining diagnosis cardiac muscle stalk Dead AUC is 0.803.Although the result of Combining diagnosis shows that three's AUC is higher, difference miRNA directly joins It is better that conjunction has, such as miR-1262 and miR-942-5p, and worse on the contrary, wherein miR-1262 and the miR-942-5p having Combining diagnosis effect close to 0.805, show very high accuracy rate.

The collection of the sample of embodiment 4 and the extraction of total serum IgE

1 sample collection：

36 myocardial infarction patients and 22 normal healthy controls crowd peripheral bloods (acquisition time in January, 2014-2015 year 8 Month).

2 Total RNAs extractions：

The processing for removing Rnase of related experiment article：

1. it will be rinsed before the application of all glasswares with DEPC and invade bubble, 120 DEG C of high pressure 20min, 180 DEG C of high temperature dry 2 More than hour.

2. by plastic ware (such as：EP pipes/pipette tips) need before use to be stayed overnight with 0.1%DEPC water enchroachment (invasion)s bubble, after drain liquid, 120 DEG C of high pressure 20min, baking box is dried standby.

Leucocyte is separated

(1) 2m1 anticoagulation cirumferential bloods are taken (blood sampling time is no more than 3h)；

(2) isometric sterile PB S are added to be sufficiently mixed in peripheral blood, cell suspension is formed；

(3) 4m1 lymphocyte separation mediums are added in another centrifuge tube；

(4) draw 4m1 cell suspensions be gently added to along tube wall lymphocyte separation medium surface (note not with lymphocyte Separating liquid is mixed).Centrifuge 1500rpm 20min；

(5) boundary layer (tunica albuginea) is gently suctioned out with suction pipe to enter in another centrifuge tube.Sterile cold PBS is washed 2 times, is washed for last 1 time Washing can move into cell suspension in EP pipes, and supernatant is removed in centrifugation, for extracting RNA.

RNA is extracted

(1) sample is taken out in -80 DEG C of refrigerators, is shredded, Trizol is added in EP pipes in 1ml/50-100mg ratio, Homogenized is carried out, 5-l0min is stored at room temperature；

(2) every milliliter of Trizol adds 0.2m1 chloroforms, acutely shakes 15s, is stored at room temperature 2-3min, 1 2000 at 4 DEG C Leave heart 15min；

(3) the careful supernatant water 600ul that makes an appointment that suctions out moves into another centrifuge tube (being careful not to be extracted into albumin layer), adds equivalent Isopropanol, overturns and mixes, be stored at room temperature 10min；

(4) 4 DEG C of 12000g centrifuge 10min, abandon supernatant, bottom visible white material；

(5) the rotation washing of the cold ethanol of 1ml 75% is added, isopropanol is cleaned；

(6) 4 DEG C of 12000g centrifuge 5min, remove the 5-l0min that dried in the air after ethanol, translucent, are dissolved with 20u1DEPC water RNA.3u1RNA samples are taken, the electrophoresis in 1.5% Ago-Gel；1u1RNA samples in UV spectrophotometer measuring concentration, RNA samples in 1.8-2.0 are considered as with A260/280 qualified.

The RT-PCR of embodiment 5 detects miR-942-5p and/or miR-1262 expression

1 RT-PCR detects miR-942-5p and/or miR-1262 expression

Reverse transcription

The preparation of RT systems：1 μ g total serum IgEs are used as template ribonucleic acid；5×miScript HiSpec Buffer 4μl；10× Nucleics Mix 2μl；miScript Reverse Transcriptase Mix 2μl；Aqua sterilisa filling-in is to 20 μ l.ABI After 37 DEG C of insulation 60min make reverse transcription reaction complete in 9700 type PCR instruments, 95 DEG C of 5min terminating reactions.Add 80 μ l Nuclease-free H₂O is diluted to 100 μ l, and to be stored in -20 DEG C of refrigerators standby.

Quantitative fluorescent PCR

The preparation of miRNA RT-PCR systems：

MiR-942-5p microRNA-specific primer primers are SEQ ID NO 3, miR-1262's MicroRNA-specific primer primers are SEQ ID NO 4.

Reaction system：MiRNAs detection of expression sets 3 parallel tube reactions every time, and internal reference is used as using snRNA U6.Expand Increasing program：95℃10min；40 circulations (95 DEG C of 10s, 60 DEG C of 30s).

3 statistical analysis

Real-time quantitative PCR sample amplified production solubility curve is all unimodal, illustrates that amplified production only has one, is specificity Amplification；Amplification curve flex point understands that amplification curve entirety collimation is good, shows that the amplification efficiency of each reaction tube is close, and the limit is put down And without raising up now, exponent phase slope is larger, illustrates that amplification efficiency is higher；According to qRT-PCR relative quantification formula meter Experimental group and the multiple proportion of control group destination gene expression are calculated, compares miR-942-5p and/or miR-1262 in myocardial infarction Expression in group and control group.As a result show：MiR-942-5p is about 2.2 times of control group in the expression quantity of illness group, MiR-1262 is at about 2.1 times that the expression quantity of illness group is control group, and result above demonstrates high-flux sequence data analysis As a result.In 58 samples, the independent Detection accuracy of miR-942-5p, miR-1262 is 84.5% and 82.8%, the two joint inspection It is 91.4% to survey accuracy rate.Display miR-942-5p, miR-1262 individually or can merge detection for myocardial infarction disease.

Although describing the present invention with reference to various preferred embodiments, it will be appreciated by those skilled in the art that can carry out each Change is planted, and available equivalents substitute base region of its component without departing from the present invention.Come in addition, many changes can be carried out Particular case or material is set to be suitable for the teachings of the present invention without departing from its base region.

SEQUENCE LISTING

<110>Beijing Yang Shen biology information technologies Co., Ltd

<120>MiRNA, composition and its application in diagnosing the illness

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Claims (10)

1. detecting application of the miRNA preparation in diagnosing myocardial infarction reagent is prepared, the miRNA is selected from one of the following Or it is several：miR-942-5p、miR-1262.

2. application according to claim 1, it is characterised in that miRNA is miR-942-5p and miR-1262 joint.

3. application according to claim 1, it is characterised in that miR-942-5p sequence is shown in sequence table SEQ ID NO 1； MiR-1262 sequence is shown in sequence table SEQ ID NO 2.

4. application according to claim 1, it is characterised in that diagnosing myocardial infarction reagent includes being based on high-flux sequence side Method and/or based on quantifying PCR method and/or based on probing procedure detection sample in miR-942-5p and/or miR-1262.

5. application according to claim 1, it is characterised in that using northern hybridizing methods, miRNA express spectra cores MiR-942-5p in piece, ribozyme protection analytical technology, RAKE methods, in situ hybridization, bead-based flow-cytometry detection sample And/or miR-1262 transcription.

6. application according to claim 1, it is characterised in that sample is peripheral blood.

7. application according to claim 1, it is characterised in that miRNA is before miR-942-5p and/or miR-1262 Body, precursor sequence is SEQ ID NO 5 and SEQ ID NO 6.

8. application according to claim 1, it is characterised in that specific amplification miR- is included based on quantifying PCR method 942-5p and/or miR-1262 primer, it is preferred that specific amplification miR-942-5p primer sequence is SEQ ID NO 3, Specific amplification miR-1262 primer sequence is SEQ ID NO 4；Based on probing procedure include with miR-942-5p and/ Or the probe of miR-1262 nucleic acid array hybridizing.

9. lower miR-942-5p and/or miR-1262 transcription and/or suppress miR-942-5p and/or miR-1262 activity Reagent prepare prevent and treat myocardial infarction preparation in application.

10. application according to claim 9, it is characterised in that using ASON, antagomiRs, miRNA sea The method of silk floss, miRNA Erasers, Target Masking and/or Mutiple Targets ASON lower miR-942-5p and/ Or miR-1262 transcription and/or blocking miR-942-5p and/or miR-1262 activity.