CN108165623A - Application of miRNA, product using miRNA and detection method - Google Patents
Application of miRNA, product using miRNA and detection method Download PDFInfo
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Abstract
The invention provides an application of miRNA, a product using the same and a detection method, and relates to the technical field of medical molecular biology, wherein a miR-17-5p, miR-126-5p or miR-145-3p marker group provided by the invention is used as a marker for auxiliary early diagnosis of heart diseases, and a mature body of the marker group stably exists in cytoplasm, so that miRNA in acute myocardial infarction serum is released into a circulating blood system earlier than a structural protein substance, and the detected level can be used as a biomarker for auxiliary early diagnosis of acute myocardial infarction. The screened three markers have strong complementarity, the three markers can be used as markers for early diagnosis of heart diseases, and the three markers have higher accuracy after being combined, so that the sensitivity and the specificity of disease diagnosis are greatly improved.
Description
Technical field
The present invention relates to Medical Molecular Biology technical field, more particularly, to a kind of application of miRNA, the production using it
Product and detection method.
Background technology
Acute myocardial infarction AMI (AMI) is that coronary artery is acute, the myocardial necrosis caused by duration hypoxic-ischemic, is the world
In the range of morbidity and mortality the main reason for one of, seriously endangered human health, according to WHO count acute myocardial infarction AMI
The primary cause of death of global range is become.The incidence of China's acute myocardial infarction AMI is sent out in recent years 55/,100,000 or so
Sick rate is on the rise.Acute myocardial infarction AMI be it is a kind of be related to Different types of etiopathogenises and a variety of physiology, biochemistry and pathological change complexity,
Critical illness.The method of the early stages reperfusion as treatment acute myocardial infarction AMI such as acute thrombolysis or early stage intervention is in constantly improve at present
And progress, unprecedented height has been reached to the treatment of acute myocardial infarction AMI, has given treatment to the same of acute myocardial infarction AMI clinical symptoms
When, people are also actively probing into acute myocardial infarction AMI molecule, gene direction, and suitable biomarker can be that Acute myocardial obstructs
Dead molecule diagnosis provides important clue and foundation, promotes early diagnosis and the realization of individualized treatment target.
In early days, acute myocardial infarction AMI is accurately diagnosed further to damage protection heart function and reduction cardiac muscle, save life
It is most important.Current traditional early diagnosis of acute myocardial infarction marker such as creatine kinase (CK), creatine kinase isozyme
(CK-MB), serum myoglobin (MYO), cardiac troponin (cTn) etc. are usually used to the early stage as acute myocardial infarction AMI
Diagnosis.Wherein, Troponin I/T is " goldstandard " of diagnosis acute myocardial infarction AMI generally acknowledged at present, is that Present clinical is generally acknowledged
Patients of acute myocardial infarction early diagnoses best marker.Patients of acute myocardial infarction is after myocardial cell injury, cell
The integrality of film, permeability change, and the chemical substance in infarct location cardiac muscle cell can constantly be discharged into peripheral blood
In, these myocardial injury markers such as CK-MB, MYO and cTn just derive from this.However, serum cTnI/cTnT is not fully
It is the specific marker object of acute myocardial infarction AMI, such as in serious acute ishemic stroke, chronic renal failure and infectivity
Shock etc. has expressed, therefore these traditional diagnostic markers there is no method to fully meet morning of acute myocardial infarction AMI
Phase diagnoses.
MicroRNA (microRNA, miRNA) is intracellular a kind of highly conserved in evolution, and length is about 22 nucleotide
Non-coding RNA gene outcome.MiRNA exists with single stranded form, by the complementary pairing with said target mrna in post-transcriptional level
Gene expression is regulated and controled, participation includes the various biologicals such as cell Proliferation, apoptosis, cell differentiation, development and Stress response
Process, therefore the occurrence and development of the numerous vital movements and disease of human body are all inseparable with the variation of miRNA.In recent years research
Further confirm that miRNA not only adjusts organism physiology pathogenesis in the cell, but also is also present in body fluid, such as serum, blood
Slurry etc., and these microRNA property stabilization, rich content, be easy to quantify detection, and there are disease specifics.It is same in this
When, in the development of genomics and proteomics, particularly chip gene expression profile and realtime quantitative inspection
(PCR) application of technology promotes many researchs about new miRNA molecule with potential clinical value.Closely
Nian Lai, many researchs find there is interwoveness from Peripheral Circulation miRNA expressions and a series of angiocardiopathies
Relationship.Such as Mamoru Satoh carry out biopsy to dilated cardiomyopathy (DCM) the patient internal membrane of heart, find and normal person's faciation
Than the horizontal significantly up-regulations of DCM patient's internal membrane of heart miR-208, long-time follow-up confirms that miR-208 is related to the Development process of DCM
Property is strong.Tijsen etc. picks out 16 kinds in the specifically expressed miRNA of patients with heart failure, evaluation then initially with miRNA chips
In show biomarker that miR-423 can be as heart failure etc..
However, up to the present, Peripheral Circulation miRNA is used for application report in early diagnosis of acute myocardial infarction very
Few, existing early diagnosis of acute myocardial infarction marker for serum miRNA in disease without expressing the characteristics of changing.
In view of this, it is special to propose the present invention.
Invention content
The present invention first be designed to provide miR-17-5p, miR-126-5p or miR-145-3p any one or
Application of many kinds of substance in the product for Diagnosing Cardiac disease is prepared, it is in the prior art to heart disease to alleviate
The technical issues of relevant specific tiny RNA research still has some deficits.
Second object of the present invention is to provide a kind of reagent for Diagnosing Cardiac disease.
Third object of the present invention is to provide a kind of kit for Diagnosing Cardiac disease.
Fourth object of the present invention be to provide miR-17-5p, miR-126-5p or miR-145-3p any one or
The detection method of many kinds of substance is asked with alleviating not strong enough the technology of the detection specificity to above-mentioned three kinds of tiny RNAs in the prior art
Topic.
The present invention provides following any one or more substances of (a)-(c) to prepare the product for Diagnosing Cardiac disease
In application：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
Further, the nucleotide sequence of the miR-17-5p is as shown in SEQ ID No.1, the miR-126-5p's
Nucleotide sequence is as shown in SEQ ID No.2, and the nucleotide sequence of the miR-145-3p is as shown in SEQ ID No.3.
Further, the product is reagent or kit.
Further, the heart disease is myocardial infarction, preferably acute myocardial infarction AMI.
The present invention also provides a kind of reagent for Diagnosing Cardiac disease, it is arbitrary that the reagent includes following (A)-(C)
One or more substances：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
The present invention also provides a kind of kit for Diagnosing Cardiac disease, the kit includes following (A)-(C)
Reagent described in any one or more substance or claim 5：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
Further, the kit further includes reverse transcriptase, buffer solution, dNTPs, MgCl2, DEPC water and Taq enzyme, with
And standard items and/or reference substance.
Further,
It is described that there is the sense primer of sequence and such as SEQ as shown in SEQ ID No.4 for detecting the primer of miR-17-5p
The downstream primer of sequence shown in ID No.5；
The primer for being used to detect miR-126-5p is with the sense primer of sequence as shown in SEQ ID No.6 and such as
The downstream primer of sequence shown in SEQ ID No.7；
The primer for being used to detect miR-145-3p is with the sense primer of sequence as shown in SEQ ID No.8 and such as
The downstream primer of sequence shown in SEQ ID No.9.
Further, the heart disease is myocardial infarction, preferably acute myocardial infarction AMI.
In addition, the present invention also provides a kind of detection method of any one or more substance of following (a)-(c), detected
The method of stating includes：It extracts sample rna and carries out fluorescent quantitation qPCR reactions, obtain any one or more substance of (a)-(c)
Relative expression quantity；
(a)-(c) be：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
MiR-17-5p, miR-126-5p or miR-145-3p marker group provided by the invention is assisted as heart disease
The marker of early diagnosis is advantageous in that miRNA only has 18-25 nucleotide in itself, and ripe body is stable in the presence of carefully
In cytoplasm, therefore miRNA very likely enters circulating earlier than structural proteins class substance release in acute myocardial infarction AMI serum
In liquid system, the biomarker of early diagnosis can be assisted as acute myocardial infarction AMI by detecting its level.Also, it filters out
Tri- markers of miR-17-5p, miR-126-5p or miR-145-3p are with strong complementarity, and three markers can be used as heart disease
Early diagnosis marker, and accuracy higher after this three markers combination, so as to greatly improve the sensitivity of medical diagnosis on disease
Property and specificity.With reference to serum miR-17-5p, miR-126-5p and miR-145-3p of unconventionality expression in heart disease, and grind
Corresponding heart disease auxiliary diagnostic box is made, is centainly had to the early diagnosis present situation of China's acute myocardial infarction AMI good
Facilitation.
Description of the drawings
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution of the prior art
Embodiment or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, in being described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, can also be obtained according to these attached drawings other attached drawings.
Fig. 1 be the miR-17-5p that provides of the embodiment of the present invention 3 patients of acute myocardial infarction PCI it is preoperative, postoperative group and just
The result figure of normal healthy control group differential expression；
Fig. 2 be the miR-126-5p that provides of the embodiment of the present invention 3 patients of acute myocardial infarction PCI it is preoperative, postoperative group and
The result figure of normal health control group differential expression；
Fig. 3 be the miR-145-3p that provides of the embodiment of the present invention 3 patients of acute myocardial infarction PCI it is preoperative, postoperative group and
The result figure of normal health control group differential expression；
Fig. 4 A are the miR-17-5p that provides of the embodiment of the present invention 3 in correlation table preoperative patients of acute myocardial infarction PCI
The result figure reached；
Fig. 4 B are the miR-17-5p that provides of the embodiment of the present invention 3 in correlation table postoperative patients of acute myocardial infarction PCI
The result figure reached；
Fig. 5 A are the miR-126-5p that provides of the embodiment of the present invention 3 in correlation preoperative patients of acute myocardial infarction PCI
The result figure of expression；
Fig. 5 B are the miR-126-5p that provides of the embodiment of the present invention 3 in correlation postoperative patients of acute myocardial infarction PCI
The result figure of expression；
Fig. 6 A are the miR-145-3p that provides of the embodiment of the present invention 3 in correlation preoperative patients of acute myocardial infarction PCI
The result figure of expression；
Fig. 6 B are the miR-145-3p that provides of the embodiment of the present invention 3 in correlation postoperative patients of acute myocardial infarction PCI
The result figure of expression；
Fig. 7 A are that miR-17-5p, miR-126-5p and miR-145-3p that the embodiment of the present invention 3 provides obstruct in Acute myocardial
The result figure of dependency expression preoperative dead patient PCI；
Fig. 7 B are that miR-17-5p, miR-126-5p and miR-145-3p that the embodiment of the present invention 3 provides obstruct in Acute myocardial
The result figure of dependency expression postoperative dead patient PCI.
Specific embodiment
Technical scheme of the present invention is clearly and completely described below in conjunction with embodiment, it is clear that described reality
It is part of the embodiment of the present invention to apply example, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
Art personnel all other embodiments obtained without making creative work belong to the model that the present invention protects
It encloses.
The present invention provides following any one or more substances of (a)-(c) to prepare the product for Diagnosing Cardiac disease
In application：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
Since miRNA only has 18-25 nucleotide in itself, ripe body is stable in the presence of in cytoplasm, change earlier than
The change of gene and albumen also earlier than the appearance of disease symptoms, thus detects the dynamic change of Peripheral Circulation miRNA, having can
It can give a clue, and then clinic can be instructed to be early diagnosed for the occurrence and development of heart disease, effectively control the hair of disease
Exhibition.Method by the use of Peripheral Circulation miRNA as early diagnosis of acute myocardial infarction molecular marked compound is than traditional special egg
White molecular detecting method will be more efficient.
In one preferred embodiment, pass through such as lower section provided by the present invention for the miRNA of Diagnosing Cardiac disease
Method screens to obtain：
(1) serum miRNA differential expressions spectrum analysis：Acute myocardial infarction AMI case and matched normal healthy controls are selected,
The preoperative cases of acute myocardial infarction AMI PCI and the matched postoperative case-controls of acute myocardial infarction AMI PCI are selected, detects its serum
MiRNA express spectras and content, analyze the general character and characteristic of miRNA between case and normal healthy controls, and analysis case PCI is preoperative and postoperative
The general character and characteristic of miRNA, screens specifically expressed miRNAs.
(2) screening disease associated serum miRNAs：To the quantitative analysis that the serum miRNAs screened is more optimized, really
Determine acute myocardial infarction AMI correlation miRNAs.
(3) serum miRNAs screenings and the development of diagnostic kit：It is opened according to acute myocardial infarction AMI associated serum miRNAs
The miRNAs auxiliary diagnostic boxes of hair realize the purpose of early diagnosis of acute myocardial infarction.
In one preferred embodiment, the nucleotides sequence of miR-17-5p is classified as：5’-
CAAAGUGCUUACAGUGCAGGUAG-3 ' (SEQ ID No.1), the nucleotides sequence of miR-126-5p is classified as：5’-
CAUUAUUACUUUUGGUACGCG-3 ' (SEQ ID No.2), the nucleotides sequence of miR-145-3p is classified as：5’-
GGAUUCCUGGAAAUACUGUUCU-3’(SEQ ID No.3)。
Wherein, the miR-17-5p involved in the present invention can be：With the identical sequence of sequence shown in SEQ ID NO.1
Arrange the bioactive functions piece either containing sequence shown in the sequence of sequence shown in SEQ ID NO.1 or SEQ ID NO.1
Section or SEQ ID NO.1 shown in sequence variant.Every sequence for having functional nucleotide sequence shown in SEQ ID NO.1, all should
Protection scope of the present invention is interpreted as, without should only be interpreted as and the identical sequence of sequence shown in SEQ ID NO.1.
MiR-126-5p involved in the present invention can be：With the identical sequence of sequence shown in SEQ ID NO.2 or
Person contain the bioactive functions segment of sequence shown in the sequence of sequence shown in SEQ ID NO.2 or SEQ ID NO.2 or
The variant of sequence shown in person SEQ ID NO.2.Every sequence for having functional nucleotide sequence shown in SEQ ID NO.2 all should understand that
For protection scope of the present invention, without should only be interpreted as and the identical sequence of sequence shown in SEQ ID NO.2.
MiR-145-3p involved in the present invention can be：With the identical sequence of sequence shown in SEQ ID NO.3 or
Person contain the bioactive functions segment of sequence shown in the sequence of sequence shown in SEQ ID NO.3 or SEQ ID NO.3 or
The variant of sequence shown in person SEQ ID NO.3.Every sequence for having functional nucleotide sequence shown in SEQ ID NO.3 all should understand that
For protection scope of the present invention, without should only be interpreted as and the identical sequence of sequence shown in SEQ ID NO.3.
In one preferred embodiment, product is reagent or kit.
In one preferred embodiment, heart disease is myocardial infarction, preferably acute myocardial infarction AMI.
The present invention also provides a kind of reagent for Diagnosing Cardiac disease, including following (A)-(C) any one or it is more
Kind substance：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
The present invention also provides a kind of kit for Diagnosing Cardiac disease, including following (A)-(C) any one or
Many kinds of substance or above-mentioned reagent：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
MiR-17-5p, miR-126-5p or the miR-145-3p provided in reagent or kit provided by the invention can be with
Risk as myocardial infarction relevant disease is estimated and is detected.Mental and physical efforts are suffered from its high sensitivity, high specificity, discovery that can be early
The risk of debilitating diseases, convenient for the prevention and treatment of early stage.
In one preferred embodiment, kit further includes reverse transcriptase, buffer solution, dNTPs, MgCl2, DEPC water
And Taq enzyme and standard items and/or reference substance.
Plurality of reagents in kit needed for setting experiment can ensure, when being tested using this kit, to make
With more convenient, operation is simpler.
In one preferred embodiment, for detecting the sense primer of miR-17-5p as miR-17-5p-F, downstream
Primer is miR-17-5p-R；For detecting the sense primer of miR-126-5p as miR-126-5p-F, downstream primer miR-
126-5p-R；For detecting the sense primer of miR-145-3p as miR-145-3p-F, downstream primer miR-145-3p-R.On
The nucleotide sequence for stating primer is as shown in the table：
| Primer | Sequence (5 ' -3 ') | Serial number |
| miR-17-5p-F | CAAAGTGCTTACAGTGCAGGTAG | SEQ ID No.4 |
| miR-17-5p-R | TTTTTTTTTTTTT | SEQ ID No.5 |
| miR-126-5p-F | CATTATTACTTTTGGTACGCG | SEQ ID No.6 |
| miR-126-5p-R | TTTTTTTTTTTTT | SEQ ID No.7 |
| miR-145-3p-F | GGATTCCTGGAAATACTGTTCT | SEQ ID No.8 |
| miR-145-3p-R | TTTTTTTTTTTTT | SEQ ID No.9 |
In one preferred embodiment, heart disease is myocardial infarction, preferably acute myocardial infarction AMI.
In addition, the present invention also provides a kind of detection method of any one or more substance of following (a)-(c), including：
It extracts sample rna and carries out fluorescent quantitation qPCR reactions, obtain the relative expression quantity of any one or more substance of (a)-(c)；
Wherein (a)-(c) is：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
MiR-17-5p, miR-126-5p or the miR-145-3p provided in the present invention can be used as heart failure correlation disease
The risk of disease is estimated and is detected.Discovery that can be early suffers from heart failure the risk of disease, convenient for the prevention and treatment of early stage.
In order to contribute to it is clearer understand present disclosure, be described in detail as follows in conjunction with specific embodiment.
1 sample process of embodiment
1. receiving patients of acute myocardial infarction 29 and physical examination normal health compareing 21, extracted outside 5mL using coagulation vessel
All blood carries out two step centrifugations at 4 DEG C, finally takes upper serum in the centrifuge tube of RNase/DNase-free.
2. serum total serum IgE extracts：Using TRI Reagent BD extracting RNAs from serum：
(1) it cracks：0.75mL TRI Reagent BD+0.25mL serum, violent mixing；
(2) internal reference：The nematode cel-mir-39 of 50pM synthesis is added in into lysate, inside is provided for checking research
Control；
(3) two-phase laminated flow：Above-mentioned lysate+0.2mL chloroforms, stand 10min after violent mixing, 4 DEG C, 12000g centrifugations
10min；
(4) RNA precipitate：+ 3 μ L glycogens of aqueous layer+0.6mL isopropanols precipitate a few hours；
(5) RNA is cleaned：1mL75% ethyl alcohol, 4 DEG C, 12000g centrifugations 5min；
(6) RNA dissolves：Add in 10 μ L DEPC water dissolutions RNA.
2 Real-time PCR of embodiment are detected
Using the miRNA detection kits of Japanese precious biotech firm (Takara) (including reverse transcription reagent box, specificity
MiRNA primers and probe, fluorescence quantitative detection kit etc.) detect miR-17-5p, miR-126-5p and miR-145- in serum
The level of 3p and object of reference miRNA (cel-mir-39), using the average value of result three times (Ct values) as the Ct of specific miRNA
Value.
The standardization of 3 data of embodiment
According to miR-17-5p, miR-126-5p, miR-145-3p in the Sample serum measured and with reference to miRNA (cel-
Mir-39 Ct values).Using cel-mir-39 as reference, relative amounts of the specific miRNA in serum is acquired, in being detected with PCR
Classical 2Δ CtMode represent specific miRNA in serum level (Δ Ct for target miRNA with reference to cel-mir-39's
The difference of Ct values).As a result as shown in Fig. 1 to Fig. 3 and Fig. 4 A to Fig. 7 B.As can be seen from Figure 1 acute myocardial infarction patients PCI hands
Cycle miR-17-5p expressions after preoperative illustrate that miR-17-5p can be developed into diagnosing obviously higher than normal healthy controls
Marker.As can be seen from Figure 2 the cycle miR-126-5p expressions of acute myocardial infarction patients PCI perioperatively are bright
It is aobvious to be higher than normal healthy controls, illustrate that miR-126-5p can be developed into diagnosis marker.As can be seen from Figure 3 Acute myocardial obstructs
The cycle miR-145-3p expressions of dead patient P CI perioperatively illustrate that miR-145-3p can obviously higher than normal healthy controls
To be developed into diagnosis marker.It can be seen that cycle miR-17-5p is preoperative in PCI as diagnosis marker from Fig. 4 A
ROC curve analysis AUC value is 0.857 (p<0.001), illustrate that there is higher accuracy rate as diagnosis marker.From Fig. 4 B
It can be seen that cycle miR-17-5p is 0.913 (p in the postoperative ROC curve analysis AUC value of PCI as diagnosis marker<
0.001), illustrate that there is higher accuracy rate as diagnosis marker.It can be seen that cycle miR-126-5p conducts from Fig. 5 A
Diagnosis marker is 0.802 (p in the preoperative ROC curve analysis AUC value of PCI<0.001), illustrate have as diagnosis marker
There is higher accuracy rate.It can be seen that cycle miR-126-5p is bent in the postoperative ROC of PCI as diagnosis marker from Fig. 5 B
Line analysis AUC value is 0.847 (p<0.001), illustrate that there is higher accuracy rate as diagnosis marker.It can from Fig. 6 A
It is 0.720 (p in the preoperative ROC curve analysis AUC value of PCI to go out to recycle miR-145-3p as diagnosis marker<0.001),
Illustrate that there is higher accuracy rate as diagnosis marker.It can be seen that cycle miR-145-3p is as diagnostic markers from Fig. 6 B
Object is 0.727 (p in the postoperative ROC curve analysis AUC value of PCI<0.001), illustrate to have as diagnosis marker higher
Accuracy rate.It can be seen that miR-17-5p, miR-126-5p, miR-145-3p are used in combination as diagnosis marker from Fig. 7 A
When, it is 0.857 (p in the preoperative ROC curve analysis AUC value of PCI<0.001), illustrate to have as diagnosis marker higher
Accuracy rate.It can be seen that miR-17-5p, miR-126-5p, miR-145-3p are used in combination as diagnosis marker from Fig. 7 B
When, it is 0.921 (p in the postoperative ROC curve analysis AUC value of PCI<0.001), illustrate to have as diagnosis marker higher
Accuracy rate.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe is described in detail the present invention with reference to foregoing embodiments, it will be understood by those of ordinary skill in the art that：Its according to
Can so modify to the technical solution recorded in foregoing embodiments either to which part or all technical features into
Row equivalent replacement；And these modifications or replacement, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
SEQUENCE LISTING
<110>University Of Qingdao
<120>The application of miRNA, product and detection method using it
<160> 9
<170> PatentIn version 3.5
<210> 1
<211> 23
<212> RNA
<213>Species home sapiens（Homo sapiens）
<400> 1
caaagugcuu acagugcagg uag 23
<210> 2
<211> 21
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<213>Species home sapiens（Homo sapiens）
<400> 2
cauuauuacu uuugguacgc g 21
<210> 3
<211> 22
<212> RNA
<213>Species home sapiens（Homo sapiens）
<400> 3
ggauuccugg aaauacuguu cu 22
<210> 4
<211> 23
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<213>Artificial sequence
<400> 4
caaagtgctt acagtgcagg tag 23
<210> 5
<211> 13
<212> DNA
<213>Artificial sequence
<400> 5
tttttttttt ttt 13
<210> 6
<211> 21
<212> DNA
<213>Artificial sequence
<400> 6
cattattact tttggtacgc g 21
<210> 7
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<213>Artificial sequence
<400> 7
tttttttttt ttt 13
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<213>Artificial sequence
<400> 8
ggattcctgg aaatactgtt ct 22
<210> 9
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<212> DNA
<213>Artificial sequence
<400> 9
tttttttttt ttt 13
Claims (10)
1. application of following any one or more substance of (a)-(c) in the product for Diagnosing Cardiac disease is prepared：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
2. application according to claim 1, which is characterized in that the nucleotide sequence of the miR-17-5p such as SEQ ID
Shown in No.1, the nucleotide sequence of the miR-126-5p is as shown in SEQ ID No.2, the nucleotides sequence of the miR-145-3p
Row are as shown in SEQ ID No.3.
3. application according to claim 1, which is characterized in that the product is reagent or kit.
4. according to claim 1-3 any one of them applications, which is characterized in that the heart disease is myocardial infarction, preferably
For acute myocardial infarction AMI.
5. a kind of reagent for Diagnosing Cardiac disease, which is characterized in that the reagent include following (A)-(C) any one or
Many kinds of substance：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
6. a kind of kit for Diagnosing Cardiac disease, which is characterized in that it is any one that the kit includes following (A)-(C)
Reagent described in kind or many kinds of substance or claim 5：
(A) for detecting the primer of miR-17-5p；
(B) for detecting the primer of miR-126-5p；
(C) for detecting the primer of miR-145-3p.
7. kit according to claim 6, which is characterized in that the kit further includes reverse transcriptase, buffer solution,
DNTPs, MgCl2, DEPC water and Taq enzyme and standard items and/or reference substance.
8. the kit described in reagent according to claim 5, claim 6 or claim 7, which is characterized in that
It is described that there is the sense primer of sequence and such as SEQ ID as shown in SEQ ID No.4 for detecting the primer of miR-17-5p
The downstream primer of sequence shown in No.5；
It is described that there is the sense primer of sequence and such as SEQ ID as shown in SEQ ID No.6 for detecting the primer of miR-126-5p
The downstream primer of sequence shown in No.7；
It is described that there is the sense primer of sequence and such as SEQ ID as shown in SEQ ID No.8 for detecting the primer of miR-145-3p
The downstream primer of sequence shown in No.9.
9. the kit described in reagent according to claim 5, claim 6 or claim 7, which is characterized in that institute
Heart disease is stated as myocardial infarction, preferably acute myocardial infarction AMI.
10. a kind of detection method of any one or more substance of following (a)-(c), which is characterized in that detect and state method packet
It includes：It extracts sample rna and carries out fluorescent quantitation qPCR reactions, obtain the opposite table of any one or more substance of (a)-(c)
Up to amount；
(a)-(c) be：
(a)miR-17-5p；
(b)miR-126-5p；
(c)miR-145-3p。
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| CN201810003212.3A CN108165623A (en) | 2018-01-02 | 2018-01-02 | Application of miRNA, product using miRNA and detection method |
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| CN201810003212.3A CN108165623A (en) | 2018-01-02 | 2018-01-02 | Application of miRNA, product using miRNA and detection method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108753956A (en) * | 2018-07-03 | 2018-11-06 | 北京泱深生物信息技术有限公司 | Application of the KDSR genes in preparing myocardial infarction diagnosis tool |
| CN110283901A (en) * | 2019-07-25 | 2019-09-27 | 青岛大学 | MiRNA probe compositions, Primer composition and diagnosis of coronary heart disease kit for diagnosis of coronary heart disease |
| WO2020088703A1 (en) * | 2018-10-31 | 2020-05-07 | Univerzita Karlova, 3.Lekarska Fakulta | A method of predicting a risk of cardiovascular disease for a child born from a complicated pregnancy |
| CN111518883A (en) * | 2020-04-02 | 2020-08-11 | 深圳大学 | Plasma miRNA marker for coronary heart disease diagnosis and application thereof |
| CN115418397A (en) * | 2022-04-19 | 2022-12-02 | 刘琳 | Biomarker for auxiliary diagnosis of dilated cardiomyopathy and application of biomarker |
-
2018
- 2018-01-02 CN CN201810003212.3A patent/CN108165623A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| JING CHEN等: "MiR-17-5p as circulating biomarkers for the severity of coronary atherosclerosis in coronary artery disease", 《INTERNATIONAL JOURNAL OF CARDIOLOGY》 * |
| 任佳君等: "循环miRNA-126 在冠心病发生发展中的作用", 《中国老年学杂志》 * |
| 肖亚利等: "血浆microRNA-145表达水平与急性心肌梗死的相关性研究", 《中国热带医学》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108753956A (en) * | 2018-07-03 | 2018-11-06 | 北京泱深生物信息技术有限公司 | Application of the KDSR genes in preparing myocardial infarction diagnosis tool |
| WO2020088703A1 (en) * | 2018-10-31 | 2020-05-07 | Univerzita Karlova, 3.Lekarska Fakulta | A method of predicting a risk of cardiovascular disease for a child born from a complicated pregnancy |
| CN110283901A (en) * | 2019-07-25 | 2019-09-27 | 青岛大学 | MiRNA probe compositions, Primer composition and diagnosis of coronary heart disease kit for diagnosis of coronary heart disease |
| CN111518883A (en) * | 2020-04-02 | 2020-08-11 | 深圳大学 | Plasma miRNA marker for coronary heart disease diagnosis and application thereof |
| CN111518883B (en) * | 2020-04-02 | 2022-12-30 | 深圳大学 | Plasma miRNA marker for coronary heart disease diagnosis and application thereof |
| CN115418397A (en) * | 2022-04-19 | 2022-12-02 | 刘琳 | Biomarker for auxiliary diagnosis of dilated cardiomyopathy and application of biomarker |
| CN115418397B (en) * | 2022-04-19 | 2023-09-19 | 刘琳 | Biomarker for auxiliary diagnosis of dilated cardiomyopathy and application thereof |
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Application publication date: 20180615 |