CN116875682A - PiRNA marker for diagnosing acute myocardial infarction heart injury, kit and application thereof - Google Patents
PiRNA marker for diagnosing acute myocardial infarction heart injury, kit and application thereof Download PDFInfo
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Abstract
The invention discloses an application of a substance for detecting a piRNA marker, which comprises one or more of the following applications: a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury; a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage; a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage; a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury; a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases; the piRNA marker is piR-hsa-28877, and the nucleotide sequence is shown as SEQ ID No. 1. piR-hsa-28877 has significantly higher expression level in the plasma of patients with acute myocardial infarction than healthy controls, indicating that it is a potential biomarker for acute myocardial infarction. The ROC curve of piR-hsa-28877 efficacy in diagnosing patients with acute myocardial infarction indicates that it has good diagnostic efficacy.
Description
Technical Field
The invention relates to a marker for diagnosing acute myocardial infarction and myocardial injury, in particular to a piRNA marker, and also relates to a corresponding kit and application thereof, belonging to the field of molecular diagnosis.
Background
Cardiovascular disease is one of the leading causes of accidental death. It is estimated that 70% of myocardial infarction is due to non-ST elevation myocardial infarction (nstemii), 30% is due to ST elevation myocardial infarction (STEMI), including new cases where left bundle branches occur in Acute Myocardial Infarction (AMI). Acute myocardial infarction is one of cardiovascular and cerebrovascular diseases which seriously threatens the survival and health of human beings, and in recent years, the mortality rate caused by the acute myocardial infarction is continuously increased.
How to improve the early diagnosis ability of myocardial infarction has been an important research direction for cardiovascular diseases. In 1955 scientists found that glutamate-oxaloacetic transaminase can appear in necrotic myocardium, after which the search and study of markers of myocardial injury has never stopped. The ideal biomarker has high clinical sensitivity and specificity, and the rise after acute myocardial infarction occurs early and has long lasting abnormal time, convenient detection and short detection time. Currently, the biomarkers commonly used clinically are protein markers. For example, markers of acute myocardial infarction include cardiac troponin (cTn), creatine kinase isozymes (CK-MB), myoglobin (Myo), and the like, with troponin being the preferred marker of acute myocardial infarction. In addition, elevation occurs to varying degrees in other diseases such as myocarditis, renal failure, autoimmune diseases. However, troponin detection as a marker also has certain limitations. Troponin elevation is generally detected in peripheral blood several hours after myocardial injury and tends to miss the optimal treatment window.
PIWI interacting RNAs (piRNAs) are RNAs consisting of 21-35 nucleotides that bind to PIWI branches of the Argonaute protein (AGO protein) family to form piRNA-guided silencing complexes. Their main targets are transposable elements (transposable element, TEs), which silence after transcription and transcription. Mutations that disrupt the piRNA pathway lead to a dramatic increase in TEs expression, decreasing germ cell maturation, and thus to a sterile phenotype. In addition to their function in the reproductive system, piRNAs are also found in a wide range of somatic cells, for example in lineage specific stem cells, such as hematopoietic and intestinal cells and adult multipotent stem cells, among others. Furthermore, piRNA is of relatively high conservation and exists as a fundamental condition for biomarkers of underlying disease.
How to obtain the biomarker which has high detection sensitivity, convenient detection and short detection window period, relates to different links of occurrence and development of cardiovascular diseases, can provide new basis for myocardial infarction diagnosis and subsequent adverse cardiovascular event occurrence prediction, and is still a hot spot for research of the technicians in the field.
Disclosure of Invention
The invention aims to provide an application of a novel piRNA marker. The marker is used for preparing substances for diagnosis, screening, prognosis evaluation and distinguishing acute myocardial infarction from myocardial injury.
The invention aims to provide a kit for detecting a novel piRNA marker. The kit can be used for diagnosis, screening, prognosis evaluation and distinguishing acute myocardial infarction from myocardial injury.
The third technical problem to be solved by the invention is to provide a primer for detecting a novel piRNA marker. The primer can be used for diagnosis, screening, prognosis evaluation and distinguishing acute myocardial infarction from myocardial injury.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
according to a first aspect of embodiments of the present invention there is provided the use of a substance for detecting a piRNA marker, comprising one or more of the following:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the piRNA marker is piR-hsa-28877, and the nucleotide sequence of the piRNA marker is shown as SEQ ID No. 1.
The "product" described above may be a product that can be used to diagnose acute myocardial infarction and myocardial injury by detecting piR-hsa-28877 expression levels by RT-PCR, real-time quantitative PCR, in situ hybridization, chip or high throughput sequencing platforms.
In the various applications, the expression of the piR-hsa-28877 in the plasma sample of the heart injury patient caused by myocardial infarction is significantly up-regulated; the expression level of piR-hsa-28877 in healthy people is significantly lower than that of myocardial infarction patients.
Wherein preferably the substance is an agent for detecting the expression level of piR-hsa-28877, or specifically recognizing piR-hsa-28877, or an agent for detecting piR-hsa-28877 content.
Wherein preferably the substance is a substance for detecting piR-hsa-28877, in particular a), b) or c) as follows:
a) Primers for detection or specific recognition of piR-hsa-28877;
b) A kit comprising said a);
c) A kit comprising said a) or said b).
Wherein, preferably, the primers are an upstream primer shown in SEQ ID No.2 and a downstream primer shown in SEQ ID No. 3.
According to a second aspect of embodiments of the present invention there is provided a kit for detecting a piRNA marker comprising one or more of the following applications:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the piRNA marker is piR-hsa-28877, and the nucleotide sequence of the piRNA marker is shown as SEQ ID No. 1; the kit comprises a reagent for detecting or specifically recognizing piR-hsa-28877 or a reagent for detecting piR-hsa-28877 expression quantity.
The kit provided by the invention can be used for detecting the expression condition of the piR-hsa-28877 characteristic gene sequence shown in SEQ ID NO.1 in the peripheral blood of a subject, and judging the probability of myocardial damage of the subject caused by acute myocardial infarction according to the information of up-regulation or down-regulation of the gene expression, thereby realizing diagnosis of the acute myocardial infarction and myocardial damage.
Kits provided herein may comprise suitable packaging and instructions for use in the methods disclosed herein. Specifically, the detection kit provided by the invention is a nucleic acid detection kit, and comprises reagents required by RNA extraction and real-time fluorescence quantitative PCR (qRT-PCR). The kit may further comprise suitable buffers and polymerases. Such kits also comprise control primers and/or probes.
Wherein, preferably, the reagent for detecting or specifically recognizing piR-hsa-28877 is a specific primer, and the specific primer is an upstream primer shown as SEQ ID No.2 and a downstream primer shown as SEQ ID No. 3.
According to a third aspect of embodiments of the present invention there is provided a primer for detecting a piRNA marker, comprising one or more of the following applications:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the primer is a primer for detecting the expression quantity of piR-hsa-28877 or specifically recognizing piR-hsa-28877.
Wherein, preferably, the primers are an upstream primer shown in SEQ ID No.2 and a downstream primer shown in SEQ ID No. 3.
Compared with the prior art, the invention has the following technical effects:
(1) The piRNA piR-hsa-28877 provided by the invention can be used as a novel marker for diagnosing acute myocardial infarction. In clinical verification experiments, the piR-hsa-28877 content expression in the blood plasma of patients with acute myocardial infarction is obviously increased compared with the expression of piR-hsa-28877 in the blood plasma of healthy controls, which shows that piR-hsa-28877 is a potential biomarker for acute myocardial infarction. The ROC curve of piR-hsa-28877 efficacy in diagnosing patients with acute myocardial infarction indicates that piR-hsa-28877 has good diagnostic efficacy.
(2) In clinical experiments, the relationship between the blood plasma CK-MB and piR-hsa-28877 expression levels of patients can be seen that the CK-MB and piR-hsa-28877 expression are positively correlated; CK-MB is a marker protein of myocardial cell damage, and piR-hsa-28877 is positively correlated with CK-MB levels. These results indicate that the levels in the plasma of piR-hsa-28877 can reflect the severity of myocardial injury disease.
(3) In vitro cytology experiments, the expression level of the anoxia-treated AC16 cells piR-hsa-28877 is increased compared with that of normal control; indicating that piR-hsa-28877 levels are elevated when cells are injured by hypoxia, which is the cellular molecular basis for piR-hsa-28877 as a diagnostic biomarker for myocardial infarction in patients.
Drawings
FIG. 1 shows the measurement of piR-hsa-28877 content in plasma of patients with acute myocardial infarction by qRT-PCR;
FIG. 2 is a ROC curve of piR-hsa-28877 for diagnosing acute myocardial infarction patients;
FIG. 3 shows the relationship between the expression levels of the patient's plasma creatine kinase isozymes CK-MB and piR-hsa-28877;
FIG. 4A shows the expression level of piR-hsa-28877 in the control and knockdown groups after transfection of AC16 with piR-hsa-28877 small interfering RNA (siRNA);
FIG. 4B is a graph showing the comparison of mRNA expression levels in control and knockdown groups after transfection of AC16 with piR-hsa-28877 small interfering RNA (siRNA);
FIG. 5 is a graph showing the relationship between the expression level of knockdown piR-hsa-28877 and piR-hsa-28877 of AC16 after hypoxia treatment.
Detailed Description
The technical contents of the present invention will be described in detail with reference to the accompanying drawings and specific examples.
It should be noted that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. The experimental procedures, which do not address the specific conditions in the examples below, are generally carried out under conventional conditions or under conditions recommended by the manufacturer. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the present invention. The preferred methods and materials described herein are presented for illustrative purposes only.
According to the invention, firstly, venous blood of 50 patients in hospital for acute myocardial infarction is used for first diagnosis, arraystar Human piRNA Array of Arraytar company is used for detecting the difference of piRNA expression profiles in serum of the patients with acute myocardial infarction and serum of corresponding normal control persons, agilent Gene Spring software is used for analyzing chip results, piRNA with obvious difference of expression quantity (Fold change is more than or equal to 2.0 and P value is less than 0.05) is screened, qRT-PCR is used for measuring the content of the first 10 piRNAs of the piRNA chip results in plasma of the patients, and the first piRNAs with obvious rising are taken for subsequent experiments. Then, the inventors again verified the relationship between the expression level of piR-hsa-28877 and the diagnosis of myocardial infarction using the in vitro experimental cell level. The specific data are as follows:
the specific data are as follows:
example 1 screening and correlation study of acute myocardial infarction myocardial injury piRNA markers1. Sample and method:
1.1 clinical samples:
venous blood was collected from 50 patients hospitalized in the general free army hospital 2018-2021 for the first diagnosis of acute myocardial infarction.
1.2 method:
1.2.1 plasma extraction: blood around human body is collected by using EDTA anticoagulation blood collection tube, centrifugated for 15min at 2500g, upper plasma is taken to 2m1 sterile tube, and the sterile tube is placed in a refrigerator at-80 ℃ for freezing.
1.2.2 RNA extraction and real-time fluorescent quantitative PCR (qRT-PCR):
total RNA was extracted from AC16 using RNAsimple Total RNA Kit (DP 419, TIANGEN, beijing, china).
(1) RNA extraction
Plasma samples were extracted for tissue RNA according to the TRIZOL Reagent (Invitrogen) instructions. To the plasma was added 1ml TRIZOL Reagent (Invitrogen) and chloroform 200u1, shaken for 20s, and allowed to stand at room temperature for 10min: centrifugal force at 13000rpm,4℃for 15min. Carefully aspirate the supernatant, add 800u1 isopropanol, gently mix upside down, stand at-20℃for 1h, centrifuge at 13000rpm at 4℃for 15min, discard the supernatant. 1ml of 75% ethanol was added, the precipitate was gently washed, centrifuged at 13000rpm at 4℃for 5min, and the supernatant was removed and dried. Adding proper amount of enzyme-free water, dissolving at 65deg.C for 10min to detect OD value and concentration of RNA, and preserving at-80deg.C for use.
(2) cDNA synthesis by RNA reverse transcription
500ng of RNA was reverse transcribed into CDNA using the reverse transcription kit (Takara RR 037A).
(3) Reverse transcription of piRNA:
in the case of the ice-on operation, each reaction system 20u1 is as follows:
TABLE 1
(4) Reverse transcription of mRNA:
in the case of the ice-on operation, each reaction system 20u1 is as follows:
TABLE 2
The reaction procedure is: maintained at 37℃for 45min,85℃for 5min, and 4 ℃.
(5)qRT-PCR
The sequences of the primers for the piRNAs were designed according to the principles of their design.
The cDNA obtained by the reverse transcription reaction was prepared according to 1:10 dilution, qRT-PCR reactions were performed as follows.
In the case of the ice-on operation, each reaction system 20u1 is as follows:
TABLE 3 Table 3
The reaction procedure of the Real-time PCR instrument for uniformly mixing the reaction liquid is as follows:
Stage 1：95℃2min；
Stage 2：Cycle 35，94℃5s，60℃1min；
Stage 3：95℃15s，60℃1min，95℃5s；
the relative levels of each mRNA were quantified with GAPDH and expressed at a relative ratio.
1.2.3 piRNA expression chip analysis:
the difference of the piRNA expression profile in the serum of acute myocardial infarction patients and the serum of corresponding normal control persons is detected by using Arraystar Human piRNA Array of Arraystar company, chip results are analyzed by using Agilent Gene Spring software, piRNA with obvious difference of expression quantity (Fold change is more than or equal to 2.0 and P value is less than 0.05) is screened, the content of the piRNA at the first 10 positions of the piRNA chip results in the plasma of the patients is detected by using qRT-PCR, and the first position with obvious increase is taken for subsequent experiments.
1.2.4 ROC curve drawing:
the subject's working characteristics (receiver operator characteristic curve, abbreviated ROC) curve is a curve plotted with true and false positive rates, and can be used to reflect the relationship between sensitivity and specificity. The sensitivity and the specificity are calculated respectively according to a series of cut-off values divided by measurement values of a test group and a control group by taking the sensitivity as an ordinate and the 1-specificity as an abscissa, and the given points are connected into a line, namely an ROC curve. ROC curves were plotted using GraphPad version 9.1.0. ROC curves reflect the diagnostic efficacy of lesion markers on disease.
1.2.5 human plasma CK-MB assay:
the detection is carried out by adopting a detection reagent of the human creatine kinase isoenzyme (CK-MB) EILSA of the Sitang organism. Firstly, 100ul of standard substances or samples to be tested are added into each hole, and the reaction plate is fully and uniformly mixed and then placed at 37 ℃ for 120 minutes. The reaction plate is fully washed for 4 to 6 times by using washing liquid, and is printed on filter paper to be dried. 100ul of primary antibody working solution was added to each well. The reaction plate was thoroughly mixed and then left at 37℃for 60 minutes. And adding 100ul of enzyme-labeled antibody working solution into each hole. The reaction plate was left at 37℃for 30 minutes. 100ul of substrate working solution is added to each hole, and the mixture is placed in a dark place at 37 ℃ for reaction for 15 minutes. 100ul of stop solution is added into each hole and mixed uniformly. Absorbance was measured at 450nm using a microplate reader over 30 minutes.
1.2.6 statistical analysis:
the t-test and analysis of variance were used for normal variables, and the Mann Whitney U test and the Kruskal Wallis test were used for non-normal variables. Statistical analysis was performed using R software (v 3.4.2) and GraphPad Prism software (v 8.00). Biological replicates are shown with individual data points superimposed on the histogram. P <0.05 was considered a significant difference.
3. Verification result:
as shown in FIG. 1, the content of piR-hsa-28877 in the plasma of patients with acute myocardial infarction is determined by qRT-PCR. The results show that the piR-hsa-28877 content expression in the blood plasma of patients with acute myocardial infarction is significantly increased compared with the piR-hsa-28877 expression in the blood plasma of healthy controls, and indicate that piR-hsa-28877 is a potential biomarker for acute myocardial infarction.
As shown in FIG. 2, the ROC curve of piR-hsa-28877 efficacy in diagnosing patients with acute myocardial infarction; the piR-hsa-28877 has good diagnosis efficacy;
as shown in FIG. 3, the relationship between the CK-MB and piR-hsa-28877 expression levels of the plasma of patients; CK-MB is a very effective index in diagnosing myocardial damage. CK-MB is mainly present in myocardial tissues, has very small content in other tissues, is a specific creatine kinase in myocardial tissues, and is a relatively sensitive index for early diagnosis of acute myocardial infarction. CK-MB in myocardial cells enters blood from cells when acute myocardial injury occurs, CK-MB in plasma is elevated, and elevation thereof has a dynamic change rule. Myocardial damage should be considered if the patient has a sequential change in the increase and decrease of CK-MB activity, and the peak value exceeds the upper limit of the reference value by a factor of 2, and no other reasons can be explained. From the figure, it can be seen that CK-MB is positively correlated with piR-hsa-28877 expression; CK-MB is a marker protein of myocardial cell damage, and piR-hsa-28877 is positively correlated with CK-MB levels. These results indicate that the levels in the plasma of piR-hsa-28877 can reflect the severity of the disease.
Example 2 in vitro experiment cell level verification of the relationship between the expression level of piR-hsa-28877 and myocardial infarction
1. Experimental materials and methods
1.1 cell culture:
human cardiomyocyte cell line (AC 16) was purchased from Gymnemonic, and cells were cultured in RPMI-1640 medium+10% foetal calf serum in a 37℃5% carbon dioxide incubator.
siRNA knockdown method: siRNA was ordered from Thermo Fisher company and, according to the instructions, added at cell confluence up to 70%, and qRT-PCR was assayed for knockdown effect after 24h induction.
1.2. Anoxic treatment:
exposing cultured cardiomyocyte AC16 cells to 95% N2/5% CO 2 The mixed gas culture chamber is subjected to anoxic treatment, and after 24 hours of anoxic treatment, the mixed gas culture chamber is subjected to anoxic treatment, and is subjected to normoxic treatment with 95% O of a control group 2 /5％CO 2 Is a cell of the heart muscle, and the level of piRNA in the cell is compared.
1.3 The qRT-PCR method was the same as in example 1.
2. Experimental results:
FIGS. 4A and 4B show comparison of piR-hsa-28877 expression level and mRNA expression level after transfection of AC16 with piR-hsa-28877 small interfering RNA (siRNA), as shown in FIGS. 4A and 4B, in which piR-hsa-28877 expression is reduced but mRNA total amount is unchanged; indicating successful transfection, and subsequent experiments were completed on this basis.
FIG. 5 shows the expression level of knockdown piR-hsa-28877 and control AC16 after hypoxia treatment of piR-hsa-28877, and the results show that hypoxia can cause the expression of piR-hsa-28877 to be significantly increased, which is probably the cytomolecular basis of piR-hsa-28877 as a diagnosis biomarker for myocardial infarction of patients.
The implementation result shows that piR-hsa-28877 has obvious correlation with myocardial infarction and is a novel biological marker of myocardial infarction. piR-hsa-28877 is up-regulated in hypoxia-treated cardiomyocytes; the ROC curve shows that piR-hsa-28877 has good capability of diagnosing acute myocardial infarction patients; piR-hsa-28877 is shown to be probably involved in the damaging effects of hypoxia on cardiomyocytes. In conclusion, piR-hsa-28877 is expected to become a novel diagnosis biomarker and treatment target of hypoxia-induced myocardial injury.
Example 3 sequence, primer and kit composition according to the invention
The nucleotide sequence of the acute myocardial infarction myocardial injury marker piR-hsa-28877 provided by the invention is shown in SEQ ID No.1,
SEQ ID No.1：GTTTCCGTAGTGTAGTGGTCATCACGTTCGCC
the primer pair for specifically recognizing piR-hsa-28877 provided by the invention comprises an upstream primer shown in SEQ ID No.2 and a downstream primer shown in SEQ ID No. 3:
SEQ ID No.2：AACAATGTTTCCGTAGTGTAGTGGTC
SEQ ID No.3：CAGTGCAGGGTCCGAGGT
the kit provided by the invention comprises the following components:
5xPrimeScript Buffer、PrimeScriptRT Enzyme Mix I、Random 6mers、Oligo dT Primer、RNase-free H20、SYBR Premix Ex Taq II(Tli RNaseH Plus)(2x)、PCR Primer(F+R)(10uM)、ROX Reference Dye(50x)。
Claims (9)
1. use of a substance for detecting a piRNA marker, characterized by comprising one or more of the following applications:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the piRNA marker is piR-hsa-28877, and the nucleotide sequence of the piRNA marker is shown as SEQ ID No. 1.
2. The use according to claim 1, wherein:
the product is a product for diagnosing acute myocardial infarction and myocardial injury by detecting piR-hsa-28877 expression level through RT-PCR, real-time quantitative PCR, in situ hybridization, a chip or a high throughput sequencing platform.
3. The use according to claim 1, wherein:
the substance is a reagent for detecting piR-hsa-28877 expression quantity or specifically recognizing piR-hsa-28877 or detecting piR-hsa-28877 content.
4. The use according to claim 1, wherein:
the substance is used for detecting piR-hsa-28877, and concretely comprises the following a), b) or c):
a) Primers for detection or specific recognition of piR-hsa-28877;
b) A kit comprising said a);
c) A kit comprising said a) or said b).
5. The use according to claim 4, wherein:
the primers are an upstream primer shown as SEQ ID No.2 and a downstream primer shown as SEQ ID No. 3.
6. A kit for detecting a piRNA marker, comprising one or more of the following applications:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the piRNA marker is piR-hsa-28877, and the nucleotide sequence of the piRNA marker is shown as SEQ ID No. 1; the kit comprises a reagent for detecting or specifically recognizing piR-hsa-28877 or a reagent for detecting piR-hsa-28877 expression quantity.
7. The kit of claim 6, wherein:
the reagent for detecting or specifically recognizing piR-hsa-28877 is a specific primer, and the specific primer is an upstream primer shown as SEQ ID No.2 and a downstream primer shown as SEQ ID No. 3.
8. A primer for detecting a piRNA marker, comprising one or more of the following applications:
a1 Use in the preparation of a product for diagnosing acute myocardial infarction and/or myocardial injury;
a2 Use in the preparation of a product for screening for acute myocardial infarction and/or myocardial damage;
a3 Use in the manufacture of a product for assessing risk of acute myocardial infarction and/or myocardial damage;
a4 Use in the preparation of a prognostic evaluation product for acute myocardial infarction and/or myocardial injury;
a5 Use in the manufacture of a product for differentiating acute myocardial infarction and/or myocardial injury from other diseases;
the primer is used for detecting the expression quantity of piR-hsa-28877 or specifically recognizing piR-hsa-28877, the piRNA marker is piR-hsa-28877, and the nucleotide sequence of the piRNA marker is shown as SEQ ID No. 1.
9. The primer of claim 8, wherein:
the primers are an upstream primer shown as SEQ ID No.2 and a downstream primer shown as SEQ ID No. 3.
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| CN112852950A (en) * | 2021-02-23 | 2021-05-28 | 张步春 | Acute myocardial infarction biomarker and application thereof |
| WO2021237238A1 (en) * | 2020-05-19 | 2021-11-25 | Cedars-Sinai Medical Center | Exosome-derived piwi-interacting rna and methods of use thereof |
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| WO2021237238A1 (en) * | 2020-05-19 | 2021-11-25 | Cedars-Sinai Medical Center | Exosome-derived piwi-interacting rna and methods of use thereof |
| CN112852950A (en) * | 2021-02-23 | 2021-05-28 | 张步春 | Acute myocardial infarction biomarker and application thereof |
| WO2022231449A1 (en) * | 2021-04-28 | 2022-11-03 | Qatar Foundation For Education, Science And Community Development | Circulating noncoding rnas as a signature of autism spectrum disorder symptomatology |
| CN115992218A (en) * | 2022-09-30 | 2023-04-21 | 中国人民解放军总医院第二医学中心 | Exosome annular RNA marker for diagnosing acute myocardial infarction, kit and application thereof |
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