CN117187379A - Application of has-miR-500b-5p in preparation of product for assisting in diagnosis of toxic myocarditis - Google Patents
Application of has-miR-500b-5p in preparation of product for assisting in diagnosis of toxic myocarditis Download PDFInfo
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- CN117187379A CN117187379A CN202311198740.6A CN202311198740A CN117187379A CN 117187379 A CN117187379 A CN 117187379A CN 202311198740 A CN202311198740 A CN 202311198740A CN 117187379 A CN117187379 A CN 117187379A
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- 238000003745 diagnosis Methods 0.000 title claims abstract description 29
- 208000009525 Myocarditis Diseases 0.000 title claims abstract description 25
- 201000008460 toxic myocarditis Diseases 0.000 title claims abstract description 20
- 210000001808 exosome Anatomy 0.000 claims abstract description 20
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 239000003550 marker Substances 0.000 claims description 17
- 230000027455 binding Effects 0.000 claims description 16
- 239000002299 complementary DNA Substances 0.000 claims description 13
- 102000044126 RNA-Binding Proteins Human genes 0.000 claims description 12
- 230000000295 complement effect Effects 0.000 claims description 12
- 239000012634 fragment Substances 0.000 claims description 12
- 229920002521 macromolecule Polymers 0.000 claims description 12
- 108700020471 RNA-Binding Proteins Proteins 0.000 claims description 10
- 239000003124 biologic agent Substances 0.000 claims description 8
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 4
- 206010047470 viral myocarditis Diseases 0.000 abstract description 18
- 108091070501 miRNA Proteins 0.000 description 13
- 239000002679 microRNA Substances 0.000 description 11
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Abstract
The application discloses an application of has-miR-500b-5p from plasma exosomes in preparing an auxiliary diagnosis toxic myocarditis product, wherein the nucleotide sequence of has-miR-500b-5p is shown in SEQ ID NO:1 is shown in the specification; according to the application, the expression quantity of has-miR-500b-5p in the plasma exosome of patients with viral myocarditis and healthy control groups is detected, and the expression quantity of has-miR-500b-5p is obviously reduced, so that the auxiliary diagnosis of viral myocarditis can be realized by detecting the expression quantity of has-miR-548a-3 p.
Description
Technical Field
The application relates to the technical field of biology, in particular to application of has-miR-500b-5p in preparation of a product for auxiliary diagnosis of toxic myocarditis.
Background
Myocarditis is an inflammatory disease of cardiac muscle, virus infection is the most common cause, clinical manifestations are different, mild cases are often manifested as chest distress, chest pain, palpitation and other discomforts, serious cases can cause cardiac insufficiency and arrhythmia, and severe myocarditis can cause cardiogenic shock and even sudden death. The death rate of the severe myocarditis is high and is about 70-80 percent. Severe viral myocarditis is extremely high in mortality rate, and the disease course can be as long as one month or even years. The disease is rapid, if the treatment is not in time, the treatment effect is poor and the prognosis is poor.
However, the clinical diagnosis of viral myocarditis is difficult, and the traditional diagnosis method comprises medical history, electrocardiogram, serum markers, heart ultrasound and heart nuclear magnetism lack sensitivity and specificity; the diagnosis needs to be carried out on endocardial myocardial biopsy, but the clinical application is severely limited due to the originality of the biopsy. Therefore, there is a need to find a highly specific, sensitive and noninvasive method for detecting viral myocarditis.
The free miRNA in the circulation has less and unstable content and is easy to be decomposed by ribonuclease, and the prospect of the free miRNA as a stable diagnosis marker and a treatment means is not clear; however, exosomes are extracellular vesicles, consisting of lipid bilayers, containing multiple protein and nucleic acid components inside, secreted by almost all cells in the body and present in multiple bodily fluids throughout the body; the exosome lipid bilayer can protect internal proteins and nucleic acids from degradation and exist stably in circulation; therefore, if the diagnosis kit for viral myocarditis is designed according to the exosome miRNA, the diagnosis and treatment level of the myocarditis can be promoted, and a new idea is provided for further research of the myocarditis.
In view of this, the present application has been made.
Disclosure of Invention
The application aims to provide an application of has-miR-500b-5p from plasma exosomes in preparing a product for assisting in diagnosing toxic myocarditis, and the expression amount of has-miR-500b-5p in the plasma exosomes of patients with viral myocarditis is obviously reduced, so that the assisting diagnosis of viral myocarditis can be realized by detecting the has-miR-500b-5 p.
In order to achieve the above object of the present application, the following technical solutions are specifically adopted:
the first aspect of the application provides application of has-miR-500b-5p from plasma exosomes in preparation of auxiliary diagnosis of toxic myocarditis products, wherein the nucleotide sequence of has-miR-500b-5p is shown in SEQ ID NO: 1.
Preferably, the product is a biological agent or a kit.
In a second aspect, the application provides a kit for aiding in the diagnosis of toxic myocarditis, comprising a marker that recognizes the has-miR-500b-5 p.
Preferably, the marker is a binding primer of cDNA complementary to the has-miR-500b-5p or a biological macromolecule binding to the has-miR-500b-5 p.
Preferably, the biological macromolecules include antibodies, functional fragments of antibodies, RNA binding proteins, and functional fragments of RNA binding proteins.
Preferably, the primer sequence of the binding primer of the cDNA complementary to the has-miR-500b-5p is shown in SEQ ID NO: 2.
In a third aspect, the application provides a biological agent for aiding in the diagnosis of toxic myocarditis, comprising a marker that recognizes the has-miR-500b-5 p.
Preferably, the marker is a binding primer of cDNA complementary to the has-miR-500b-5p or a biological macromolecule binding to the has-miR-500b-5 p.
Preferably, the biological macromolecules include antibodies, functional fragments of antibodies, RNA binding proteins, and functional fragments of RNA binding proteins.
Preferably, the primer sequence of the binding primer of the cDNA complementary to the has-miR-500b-5p is shown in SEQ ID NO: 2.
Compared with the prior art, the application has the beneficial effects that at least:
according to the application, the expression level of has-miR-500b-5p in the plasma exosomes of patients with viral myocarditis and healthy control groups is detected, and the expression level of has-miR-500b-5p is found to be obviously reduced, so that the auxiliary diagnosis of viral myocarditis can be realized by detecting the expression level of has-miR-548a-3 p.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. Like elements or portions are generally identified by like reference numerals throughout the several figures. In the drawings, elements or portions thereof are not necessarily drawn to scale.
FIG. 1 shows the expression differences of exosome miRNAs obtained by RNA sequencing in example 1 of the present application;
FIG. 2 is the result of qRT-PCR verification of differential expression of 6 miRNAs in example 2 of the present application.
Detailed Description
Embodiments of the technical scheme of the present application will be described in detail below with reference to the embodiments. The following examples are only for more clearly illustrating the technical aspects of the present application, and thus are merely examples, and are not intended to limit the scope of the present application.
It is noted that unless otherwise indicated, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this application belongs.
The embodiment of the application provides an application of has-miR-500b-5p in preparing a product for assisting in diagnosing toxic myocarditis, wherein the nucleotide sequence of has-miR-500b-5p is shown in SEQ ID NO:1, specifically, the sequence from the 5 'end to the 3' end is: AAUCCUUGCUACCUGGGU.
In the present application, the types of the above products are not particularly limited, and those skilled in the art can select according to actual needs, and in one embodiment, the above product types may be biological agents, and in other embodiments, the above product types may be kits.
Another embodiment of the application provides a kit for aiding in the diagnosis of toxic myocarditis, comprising a marker that recognizes the has-miR-500b-5 p.
The has-miR-500b-5p is identified by identifying the marker of the has-miR-500b-5p, so that the detection of the expression quantity of the has-miR-500b-5p is realized, and further, the auxiliary diagnosis of viral myocarditis can be realized according to the detection result of the expression quantity of the has-miR-500b-5 p.
In one embodiment, the marker may be a binding primer for the cDNA complementary to the has-miR-500b-5 p; in other embodiments, the marker may be a biological macromolecule that binds to the has-miR-500b-5 p.
Further, the biological macromolecules can be antibodies, antibody functional fragments, RNA binding proteins, and RNA binding protein functional fragments.
In order to better realize the detection of the has-miR-500b-5p, in one embodiment, the primer sequence of the binding primer of the cDNA complementary to the has-miR-500b-5p is shown in SEQ ID NO:2, in particular GAATCCTTGCTACCTGGGTAAA.
Yet another embodiment of the present application provides a biological agent for aiding in the diagnosis of toxic myocarditis comprising a marker that recognizes the has-miR-500b-5 p.
The has-miR-500b-5p is identified by identifying the marker of the has-miR-500b-5p, so that the detection of the expression quantity of the has-miR-500b-5p is realized, and further, the auxiliary diagnosis of viral myocarditis can be realized according to the detection result of the expression quantity of the has-miR-500b-5 p.
In one embodiment, the marker may be a binding primer for the cDNA complementary to the has-miR-500b-5 p; in other embodiments, the marker may be a biological macromolecule that binds to the has-miR-500b-5 p.
Further, the biological macromolecules can be antibodies, antibody functional fragments, RNA binding proteins, and RNA binding protein functional fragments.
To better enable detection of has-miR-500b-5p in plasma exosomes, in one embodiment, the upstream primer sequence of the binding primer of the cDNA complementary to has-miR-500b-5p is set forth in SEQ ID NO:2, in particular GAATCCTTGCTACCTGGGTAAA.
The technical scheme of the application is further described in detail through specific examples.
Example 1
The embodiment adopts a high-throughput sequencing technology to detect the expression quantity of miRNA in the plasma exosomes of patients with viral myocarditis and a control group (normal people);
the detection result is shown in figure 1;
as can be seen from fig. 1:
there was a significant difference in exosome mirnas between the two groups, with a total of 28 differentially expressed mirnas, 14 of which were up-regulated and 14 of which were down-regulated.
Example 2
The present example is qRT-PCR detection of miRNA expression in plasma exosomes of viral myocarditis (8 humans) and normal healthy (8 human) control group:
according to the expression difference condition, 8 miRNAs obtained in the example 1 are screened to obtain 6 miRNAs with the largest expression difference, namely has-miR-500b-5p, has-miR-208a-3p, has-miR-197-5p, has-miR-548a-3p, has-miR-6782-5p and has-miR-412-5p;
collecting peripheral blood samples of patients with viral myocarditis and healthy control groups, separating by using an ultra-high speed centrifugation method to obtain exosomes, separating miRNA in the exosomes by using a Magen HiPure Serum/Plasma miRNA Kit, reversely transcribing RNA of the two groups of samples into cDNA, and amplifying and quantifying by qRT-PCR, wherein the detection result is shown in figure 2;
as can be seen from fig. 2:
the results of the sequencing of the has-miR-500b-5p and has-miR-548a-3p in the plasma exosome of the patient with viral myocarditis are consistent with those of the embodiment 1, and the has-miR-500b-5p in the plasma exosome of the patient with viral myocarditis is obviously reduced; therefore, the purpose of auxiliary diagnosis of viral myocarditis can be achieved by specifically detecting has-miR-500b-5p in plasma exosomes.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present application, and not for limiting the same; although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the application, and are intended to be included within the scope of the appended claims and description.
Claims (10)
1. Application of has-miR-500b-5p from plasma exosomes in preparing auxiliary diagnosis toxic myocarditis products is characterized in that the nucleotide sequence of has-miR-500b-5p is shown in SEQ ID NO: 1.
2. The use according to claim 1, wherein the product is a biological agent or a kit.
3. A kit for aiding in the diagnosis of toxic myocarditis comprising a marker that recognizes the has-miR-500b-5p of claim 1.
4. The kit for aiding diagnosis of toxic myocarditis according to claim 3, wherein the marker is a binding primer of cDNA complementary to the has-miR-500b-5p or a biological macromolecule binding to the has-miR-500b-5 p.
5. The kit for aiding in the diagnosis of toxic myocarditis according to claim 4, wherein the biological macromolecules comprise antibodies, functional fragments of antibodies, RNA binding proteins and functional fragments of RNA binding proteins.
6. The kit for aiding in the diagnosis of toxic myocarditis according to claim 4, wherein the primer sequence of the binding primer of the cDNA complementary to has-miR-500b-5p is shown in SEQ ID NO: 2.
7. A biological agent for aiding in the diagnosis of toxic myocarditis comprising a marker that recognizes the has-miR-500b-5p of claim 1.
8. The biological agent for aiding diagnosis of toxic myocarditis according to claim 7, wherein the marker is a binding primer of the cDNA complementary to the has-miR-500b-5p or a biological macromolecule binding to the has-miR-500b-5 p.
9. The biologic for aiding in the diagnosis of toxic myocarditis according to claim 7, wherein the biologic macromolecules comprise antibodies, functional fragments of antibodies, RNA binding proteins and functional fragments of RNA binding proteins.
10. The biological agent for aiding diagnosis of toxic myocarditis according to claim 4, wherein the primer sequence of the binding primer of the cDNA complementary to has-miR-500b-5p is shown in SEQ ID NO: 2.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311198740.6A CN117187379A (en) | 2023-09-18 | 2023-09-18 | Application of has-miR-500b-5p in preparation of product for assisting in diagnosis of toxic myocarditis |
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| CN202311198740.6A CN117187379A (en) | 2023-09-18 | 2023-09-18 | Application of has-miR-500b-5p in preparation of product for assisting in diagnosis of toxic myocarditis |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112779329A (en) * | 2021-01-25 | 2021-05-11 | 中南大学湘雅三医院 | Molecular marker for auxiliary diagnosis of viral meningitis and application and kit thereof |
| CN115418397A (en) * | 2022-04-19 | 2022-12-02 | 刘琳 | Biomarker for auxiliary diagnosis of dilated cardiomyopathy and application of biomarker |
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- 2023-09-18 CN CN202311198740.6A patent/CN117187379A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112779329A (en) * | 2021-01-25 | 2021-05-11 | 中南大学湘雅三医院 | Molecular marker for auxiliary diagnosis of viral meningitis and application and kit thereof |
| CN115418397A (en) * | 2022-04-19 | 2022-12-02 | 刘琳 | Biomarker for auxiliary diagnosis of dilated cardiomyopathy and application of biomarker |
Non-Patent Citations (1)
| Title |
|---|
| QINCHAO WU ET AL.: ""Transcriptional and functional analysis of plasma exosomal microRNAs in acute viral myocarditis"", 《GENOMICS》, vol. 116, 30 December 2023 (2023-12-30), pages 1 - 9 * |
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