MascRNA for diagnosing, screening or evaluating acute coronary syndrome and application thereof

Abstract

The invention belongs to the technical field of biomedicine, and discloses mascRNA for diagnosing, screening or evaluating acute coronary syndrome and application of mascRNA in preparation of products for diagnosing, screening or evaluating prognosis of acute coronary syndrome. The novel marker mascRNA provided by the invention has excellent stability, and brings convenience and low trauma to diagnosis of the acute coronary syndrome; the expression difference in plasma exosomes of patients with the acute coronary syndrome and people with normal coronary arteries is achieved, and the important value is achieved in the aspect of clinical diagnosis of the acute coronary syndrome (AUC is 0.763, and 95% CI is 0.702-0.824). Kaplan-Meier analysis and log-rank test data show that compared with a patient with low mascRNA expression, a patient with high mascRNA expression has a lower MACE-free survival rate (Plt; 0.001) in the formula (1).

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and particularly relates to a mascRNA for diagnosing, screening or evaluating acute coronary syndrome and an application thereof in preparing a product for diagnosing, screening or evaluating acute coronary syndrome. Background Art

[0002] In recent years, cardiovascular diseases, mainly atherosclerotic cardiovascular disease (ASCVD), account for more than 40% of the causes of death. Among them, acute coronary syndrome (ACS) is a serious manifestation of ASCVD. It is a group of acute myocardial ischemic clinical syndromes characterized by complete or incomplete occlusive thrombosis caused by rupture or invasion of coronary atherosclerotic plaques, including ST-segment elevation myocardial infarction (STEMI), non-ST-segment elevation myocardial infarction (NSTEMI) and unstable angina (UA). The symptoms of ACS are complex and diverse, mainly manifested as paroxysmal chest pain and chest tightness. The patient's condition usually develops rapidly and changes rapidly. If effective treatment measures are not taken in time, it may cause serious complications such as arrhythmia and heart failure. Severe cases may even endanger life safety. Therefore, early diagnosis and treatment of the disease are directly related to the patient's prognosis and survival rate.

[0003] At present, the clinical diagnosis of ACS is usually based on the patient's clinical symptoms, characteristic changes in electrocardiogram, myocardial enzyme spectrum detection, and combined with coronary angiography, coronary angiography and other imaging auxiliary examinations to assist in diagnosis. Most ACS patients are diagnosed in the emergency department for the first time due to chest pain or chest tightness as the main complaint. However, some patients are difficult to diagnose early because of their untypical clinical symptoms. Coronary angiography has high value as the "gold standard" for diagnosing ACS, but it is traumatic and easily restricted by medical conditions and costs, and its flexibility and convenience are poor. Although the myocardial necrosis markers used in clinical laboratory tests, such as troponin I (cTnI), creatine kinase (CK), and creatine kinase isoenzyme (CKMB), have the characteristics of early increase and long duration of increase, the changes in the levels of myocardial necrosis markers still lag behind the progression of ACS due to the rapid progression of ACS, resulting in delayed diagnosis and missed diagnosis. Therefore, it is currently necessary to find a biomarker that can help to diagnose ACS early and quickly and can timely assess the severity of ACS.

[0004] MALAT1-associated small cytoplasmic RNA (mascRNA) is a highly conserved tRNA-like small noncoding RNA composed of 61 nucleotides, which is derived from the post-transcriptional processing of the initial transcript of lncRNAMALAT1. Studies have shown that MALAT1 is widely involved in tumor development and other cellular processes, such as regulating lipopolysaccharide-induced inflammatory responses. MascRNA has also been recently shown to have the function of promoting tumor cell proliferation and migration. Given that mascRNA is mainly highly expressed in immune cells such as monocytes / macrophages, its regulatory role in immune function has attracted much attention. Studies have reported that knockdown of mascRNA affects the expression of multiple immune genes in monocytes, while exogenous mascRNA can enhance the resistance of cardiomyocytes to viral infection. However, there are no reports showing that mascRNA is correlated with the diagnosis or prognosis of ACS. Summary of the invention

[0005] In order to overcome the shortcomings and deficiencies of the prior art, the object of the present invention is to provide a use of mascRNA (MALAT1-associated cytoplasmic small RNA) in the preparation of products for diagnosing, screening or evaluating the prognosis of acute coronary syndrome.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] In a first aspect, the present invention provides the use of mascRNA (MALAT1-associated cytoplasmic small RNA) in the preparation of any of the following products:

[0008] (1) Diagnosis of acute coronary syndrome;

[0009] (2) Screening for acute coronary syndrome;

[0010] (3) Assess the prognosis of acute coronary syndrome.

[0011] Furthermore, the product includes a reagent for detecting the expression level of mascRNA.

[0012] Furthermore, the mascRNA is specifically present in plasma.

[0013] Furthermore, the expression level of mascRNA in patients with acute coronary syndrome was significantly higher than that in those with normal coronary arteries.

[0014] Furthermore, the expression level of mascRNA in the plasma of patients with acute coronary syndrome was significantly higher than that of those with normal coronary arteries.

[0015] Furthermore, the expression level of mascRNA in plasma exosomes of patients with acute coronary syndrome was significantly higher than that of those with normal coronary arteries.

[0016] The present invention detects the expression level of mascRNA in plasma exosomes of patients with coronary syndrome and normal coronary arteries. The results show that compared with patients with normal coronary arteries, the expression of mascRNA in peripheral blood plasma exosomes of patients with acute coronary syndrome is significantly increased, and has good stability and diagnostic performance. Detecting the expression level of mascRNA has significant advantages in diagnosing acute coronary syndrome, including convenient operation, small trauma and no radiation hazard, overcoming the limitations of traditional diagnostic methods. The ROC curve shows that the AUC is 0.763 and the 95% CI is 0.702-0.824. Kaplan-Meier analysis and log-rank test data show that compared with patients with low expression of mascRNA, patients with high expression of mascRNA have a lower MACE-free survival rate (P<0.001). Cox regression analysis showed that after correcting age, diabetes and LVEF, mascRNA was significantly associated with the occurrence of 1-year MACEs, with an HR of 2.959 and a 95% CI of 1.187-4.669 (P<0.001). This shows that mascRNA has good clinical application value in the screening and prognosis assessment of acute coronary syndrome and is suitable for promotion and application in clinical practice.

[0017] Based on this, the present invention proposes for the first time a biomarker mascRNA for diagnosing, screening or evaluating the prognosis of acute coronary syndrome, provides a mascRNA-based method for diagnosing acute coronary syndrome, and proposes the use of mascRNA (MALAT1-associated cytoplasmic small RNA) in the preparation of products for diagnosing, screening or evaluating the prognosis of acute coronary syndrome.

[0018] Furthermore, the product may include at least one of a chip, a preparation, a kit, a test paper or a high-throughput sequencing platform.

[0019] Furthermore, the product can diagnose, screen or evaluate the prognosis of acute coronary syndrome by detecting the expression level of mascRNA in a sample.

[0020] In a second aspect, the present invention also provides the use of a product for detecting the expression level of mascRNA in the preparation of any of the following products:

[0021] (1) Diagnosis of acute coronary syndrome;

[0022] (2) Screening for acute coronary syndrome;

[0023] (3) Assess the prognosis of acute coronary syndrome.

[0024] Furthermore, the products for detecting the expression level of mascRNA include: products for detecting the expression level of mascRNA by RT-PCR, fluorescent quantitative PCR, immunoassay, in situ hybridization, chip or high-throughput sequencing platform.

[0025] Furthermore, the product for detecting the expression level of mascRNA by RT-PCR includes at least a pair of primers for specifically amplifying mascRNA.

[0026] Furthermore, the product for detecting the expression level of mascRNA by fluorescent quantitative PCR includes at least a pair of primers for specifically amplifying mascRNA.

[0027] Further, the sources of mascRNA and its expression products for diagnosing, screening or evaluating acute coronary syndrome include but are not limited to body fluids such as blood, tissue fluid, urine, saliva, and cerebrospinal fluid. In a specific embodiment of the present invention, the source of mascRNA and its expression products for diagnosing, screening or evaluating the prognosis of acute coronary syndrome is blood. Further, the sample is peripheral venous blood. Further, the sample is exosomes in plasma.

[0028] Furthermore, the above-mentioned product was used to detect the expression level of mascRNA in the plasma exosomes of patients with acute coronary syndrome and those with normal coronary arteries for diagnosis. The results showed that the expression level of mascRNA was significantly increased in patients with acute coronary syndrome.

[0029] In a third aspect, the present invention also provides the use of mascRNA in preparing a fluorescent quantitative PCR kit for diagnosing acute coronary syndrome.

[0030] Furthermore, the kit includes at least a pair of primers for specifically amplifying mascRNA, and the sequences are shown in SEQ ID NO.1 and SEQ ID NO.2.

[0031] Forward primer 5′-GATGCTGGTGGTTGGCACTC-3′ (SEQ ID NO. 1);

[0032] Reverse primer: 5'-TGGAGACGCCGCAGGGAT-3' (SEQ ID NO. 2).

[0033] In a fourth aspect, the present invention provides a fluorescent quantitative PCR kit for diagnosing acute coronary syndrome.

[0034] Furthermore, the kit includes at least a pair of primers for specifically amplifying mascRNA, and the sequences are shown in SEQ ID NO.1 and SEQ ID NO.2.

[0035] Furthermore, the kit also includes a pair of primers for specifically amplifying the internal reference U6, the sequences of which are shown in SEQ ID NO.3 and SEQ ID NO.4.

[0036] Forward primer 5′-CTCGCTTCGGCAGCACA-3′ (SEQ ID NO. 3);

[0037] Reverse primer: 5'-AACGCTTCACGAATTTGCGT-3' (SEQ ID NO. 4).

[0038] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0039] The present invention provides a new marker mascRNA for diagnosing acute coronary syndrome, which has excellent stability and brings convenience and low trauma to the diagnosis of acute coronary syndrome. The present invention found that mascRNA was different in the plasma exosomes of patients with acute coronary syndrome and those with normal coronary arteries, and it has important value in the clinical diagnosis of acute coronary syndrome (AUC is 0.763, 95% CI is 0.702-0.824). Kaplan-Meier analysis and log-rank test data showed that patients with high expression of mascRNA had a lower MACE-free survival rate (P<0.001) compared with patients with low expression of mascRNA. Cox regression analysis showed that after correcting age, diabetes and LVEF, mascRNA was significantly associated with the occurrence of 1-year MACEs, with HR of 2.959 and 95% CI of 1.187-4.669 (P<0.001). This shows that mascRNA has good clinical application value in the screening and prognosis assessment of acute coronary syndrome, and detecting the expression level of mascRNA has significant advantages in the diagnosis of acute coronary syndrome, including convenient operation, less trauma and no radiation hazard, overcoming the limitations of traditional diagnostic methods and being suitable for promotion and application in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0041] Figure 1 Schematic diagram of the research process of the present invention.

[0042] Figure 2 Centrifugation protocol for enrichment of plasma exosomes.

[0043] Figure 3 For the identification of exosomes. Among them, A is the transmission electron microscopy (TEM) analysis of exosome morphology; B is the Western blot analysis of exosome markers; C is the nanoparticle tracking analysis to determine the exosome particle size.

[0044] Figure 4 is the expression of mascRNA in exosomes in ACS patients. A is the expression level of mascRNA in exosomes of ACS patients and non-ACS patients; B is the expression level of mascRNA in exosomes of ACS patients with and without MACE during the 1-year follow-up.

[0045] Figure 5 The correlation between mascRNA expression level in exosomes and Gensini.

[0046] Figure 6 The diagnostic value of mascRNA expression level in exosomes for ACS.

[0047] Figure 7 The predictive value of mascRNA expression level in exosomes for the prognosis of ACS patients.

[0048] Among them, *P<0.05, **P<0.01. DETAILED DESCRIPTION

[0049] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0050] In order to facilitate the understanding of the present invention, the present invention is described more comprehensively below in conjunction with preferred embodiments and drawings. It should be understood that the specific description below is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0051] Patients with acute coronary syndrome (ACS) were diagnosed in accordance with the diagnostic criteria for ACS established by the American Heart Association (AHA) and the American College of Cardiology (ACC), with patients with chest pain but normal coronary arteries serving as controls. Exclusion criteria included: previous severe valvular heart disease, structural heart disease, severe arrhythmia, acute or chronic inflammation, malignant tumors, severe liver and kidney dysfunction, autoimmune diseases, and blood system diseases. The patient selection process can be found in Figure 1 .

[0052] Example 1: Isolation and identification of exosomal mascRNA expression in plasma of patients with acute coronary syndrome and those with normal coronary arteries

[0053] 1. Research subjects

[0054] A total of 190 subjects were included in this study, and all patient specimens were obtained from Meizhou People's Hospital. Among them, subjects with normal coronary arteries were matched with patients with coronary atherosclerosis by age, gender, hypertension, diabetes, and hyperlipidemia. Finally, 140 ACS patients and 50 control patients were matched. The study protocol was approved by the local ethics committee and conducted in accordance with the ethical standards set forth in the Declaration of Helsinki (Approval No.: 2023-C-34). The informed consent form was signed by the patient. The baseline data of the patients are shown in Table 1.

[0055] Table 1

[0056]

[0057]

[0058] Note: LVEF: left ventricular ejection fraction; TG: triglyceride; TC: total cholesterol; LDL-C: low-density lipoprotein cholesterol; HDL-C: high-density lipoprotein cholesterol; WBC: white blood cell; cTnI: cardiac troponin I; NA: not applicable.

[0059] 2. Specimen Collection

[0060] 2.1 Sample preparation: Collect venous blood from patients when they are admitted to the hospital, place it in a blood collection tube containing EDTA anticoagulant, and mix it by inversion to prevent blood coagulation;

[0061] 2.2 Plasma separation: The collected blood samples were centrifuged at 2500 g for 15 min at room temperature to remove cells and cell debris, separate the plasma, and transfer all the supernatant (i.e., plasma) to a new centrifuge tube;

[0062] 2.3 Low-speed centrifugation: Place the plasma sample at 4°C and centrifuge at 300g for 10 minutes to remove residual cells and large particles. After centrifugation, carefully transfer the supernatant to a new sterile centrifuge tube and discard the precipitate.

[0063] 2.4 Medium-speed centrifugation: Centrifuge at 2000g for 10 min at 4°C to remove dead cell debris and larger particles. After the centrifugation, place the centrifuge tube in an ice bath and slowly pour the supernatant into a new sterile centrifuge tube;

[0064] 2.5 High-speed centrifugation: Centrifuge at 10,000 g for 10 min at 4°C to further remove cell debris and larger impurity particles. After the centrifugation is completed, slowly pour the supernatant into a new sterile centrifuge tube and discard the precipitate;

[0065] 2.6 Ultracentrifugation: Centrifuge at 120,000 g for 30 min at 4°C to precipitate the exosomes, and collect the precipitate to obtain the exosomes;

[0066] 2.7 Exosome washing and resuspension: Add pre-cooled PBS to the centrifuge tube, gently vortex or tap the bottom of the tube to resuspend the exosome pellet, centrifuge the suspension at 120000g for 30min, carefully discard the supernatant after centrifugation, and resuspend the exosomes in 100μL PBS and store at -80℃ for later use (see the figure for exosome collection and separation). Figure 2 ).

[0067] 3. Exosome Extraction and Identification

[0068] Transmission electron microscopy, nanoparticle tracking analysis, and Western Blot (WB) were used to identify the isolated exosomes. Transmission electron microscopy: Exosomes showed a typical double-layer vesicle structure, which was round or quasi-round ( Figure 3 A); Western blot results showed that high abundance of exosome marker proteins CD9, TSG101 and CD63 could be detected in the isolated plasma exosomes ( Figure 3 B), while these proteins were not detected in the supernatant of exosome removal; Nanoparticle tracking analysis: the average diameter of vesicles in the sample was 130nm ( Figure 3 C).

[0069] 4. Determination of relative expression of exosomal mascRNA

[0070] Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to determine the relative expression of mascRNA in plasma exosomes: exosome RNA was extracted using the SteadyPure small RNA extraction kit (Accurate Biology, China).

[0071] 4.1 Extraction of exosome RNA: Total exosome RNA was extracted using the SteadyPure Small RNAExtraction Kit from Acori Biotech. Collect the exosome precipitate obtained by ultracentrifugation, add 1 mL of lysis solution to it, mix it with a pipette, and let it stand at room temperature for 2 minutes. Add 200 μL of chloroform to the above lysis solution, mix it thoroughly, and let it stand at room temperature for 5 minutes. Centrifuge the above centrifuge tube at 12000g and 4℃ for 15 minutes. Pipette 450 μL of supernatant, put it into a new centrifuge tube, slowly add 225 μL of anhydrous ethanol, and mix it with a pipette. Use Nano Drop 2000 to detect the concentration and purity of total RNA.

[0072] 4.2 Reverse transcription and cDNA synthesis

[0073] ① Remove PrimeScript TM RT reagent Kit (TaKaRa, Dalian, China), centrifuge briefly, and place on ice. Prepare the reverse transcription reaction solution according to the components in Table 2. The reagents are prepared on ice:

[0074] Table 2 Reverse transcription reaction system

[0075] Reagents Usage 5×PrimeScript RT Master Mix 2μL Total RNA 300ng-500ng <![CDATA[RNase Free dH 2 The]]> up to 10μL

[0076] ② After gently mixing the prepared mixture, turn on the PCR instrument, set the program according to the following cycle, put the samples into the instrument one by one, and start the reverse transcription reaction. The reverse transcription reaction program is: 37℃15min, 85℃5s. After the reaction is completed, take out the reverse transcription reaction solution, i.e. the synthesized cDNA.

[0077] 4.3 Fluorescence real-time quantitative PCR (RT-qPCR) detection

[0078] TB Green Premix Ex Taq II (Tli RNaseH Plus) was used for RT-qPCR, and U6 was used as an internal reference. The relative expression of mascRNA genes in plasma exosomes of different groups was detected by RT-qPCR reaction, and the expression differences between them were compared. The nucleotide sequences of mascRNA specific amplification primers are shown in SEQ ID NO.1 and SEQ ID NO.2. The nucleotide sequences of U6 specific amplification primers are shown in SEQ ID NO.3 and SEQ ID NO.4.

[0079] Prepare the RT-qPCR reaction system according to the ratio in Table 3. Prepare the reagents on ice:

[0080] Table 3 RT-qPCR reaction system

[0081] Reagents Usage TBGreenPremixExTaqII(TliRNaseHPlus) 10μL PCRForwardPrimer (10 μM) 1μL PCR Reverse Primer (10 μM) 1μL cDNA template 2μL <![CDATA[RNaseFreedH 2 The]]> 6μL Total 20μL

[0082] The two-step method was used to perform RT-qPCR and generate melting curves. The program settings are shown in Table 4.

[0083] Table 4 Two-step RT-qPCR reaction system program settings

[0084]

[0085] After the reaction, the relative expression of mascRNA gene was calculated by 2-ΔΔCt based on the original RT-qPCR detection results and U6 as the internal reference.

[0086] 5. ROC curve analysis and AUC (area under the curve) were used to evaluate the diagnostic efficacy of exosomal mascRNA in ACS patients. The results are shown in Figure 4 and Figure 5 .

[0087] 6. Correlation between exosomal mascRNA expression levels and the risk of ACS

[0088] The clinical coronary angiography results of all patients were collected, and the coronary stenosis score (modified Gensini score) was performed according to the coronary angiography results of patients with coronary heart disease. The scoring method was as follows: the extent and severity of proximal coronary artery lesions were scored by scoring each lesion site: less than 50% luminal diameter stenosis, 1 point; 50%-74% stenosis, 2 points; 75%-99% stenosis, 3 points; complete occlusion, 4 points. The scores of each lesion in the proximal coronary circulation were summarized to obtain the final total score of the severity of coronary atherosclerosis (Gensini score). Spearman correlation analysis was used to detect the correlation between exosomal mascRNA and Gensini score. At the same time, a multivariate logistic regression model was used to evaluate the correlation between exosomal mascRNA and the risk of ACS.

[0089] 7. Evaluation value of exosomal mascRNA in ACS patients one year after PCI

[0090] ACS patients were followed up for one year after PCI via electronic medical records or telephone follow-up. The primary outcome measure was the incidence of major adverse cardiovascular events (MACE), including all-cause death, non-fatal myocardial infarction, target vessel revascularization, rehospitalization for angina or heart failure, and stent thrombosis. Kaplan-Meier analysis and log-rank test were used to evaluate the 1-year MACEs-free survival rate in the high-mascRNA group and the low-mascRNA group. Multivariate Cox regression was used to analyze the relationship between exosomal mascRNA and MACEs in ACS patients.

[0091] 8. Results Analysis

[0092] 8.1 Expression of exosomal mascRNA in ACS patients:

[0093] Depend on Figure 4 It can be seen that compared with non-ACS patients, the expression of exosome mascRNA in ACS patients was increased ( Figure 4 A). Comparison of the exosomal mascRNA expression between patients with MACEs and those without MACEs during the 1-year follow-up after PCI treatment revealed that the masc RNA expression in the MACE group was significantly higher than that in the non-MACE group ( Figure 4 B). Further analysis of the correlation between exosomal mascRNA and clinical parameters: Spearman correlation analysis showed that the level of exosomal mascRNA was significantly positively correlated with the Gensini score (r=0.242, P<0.001) ( Figure 5 ).

[0094] 8.3 Correlation between exosomal mascRNA expression levels and the risk of ACS:

[0095] ROC curve analysis showed that exosomal mascRNA could be used as a diagnostic predictor for ACS, with an AUC of 0.763 and a 95% CI of 0.702-0.824 ( Figure 6 ). A multivariate logistic regression model was used to evaluate the correlation between exosomal mascRNA and ACS, and it was found that compared with patients in the first quartile (Q1) of mascRNA expression, patients in the second, third, and fourth quartiles had an increased risk of ACS (OR: 3.423, 95% CI: 1.427-8.213; OR: 5.542, 95% CI: 1.859-16.524; OR: 9.288, 95% CI: 3.275-26.340; P values ​​were all < 0.01; Table 5).

[0096] Table 5 Relationship between exosomal mascRNA expression and the risk of ACS

[0097]

[0098]

[0099] *Adjusted for age, sex, hypertension and diabetes.

[0100] 8.4 Predictive value of exosomal mascRNA for major cardiovascular events in ACS patients one year after PCI:

[0101] The patients were divided into a high mascRNA group (≥3.85, n=60) and a low mascRNA group (<3.85, n=60). Kaplan-Meier analysis and log-rank test were used to evaluate the 1-year MACEs-free survival rate in the high and low mascRNA groups. The data showed that patients with high mascRNA expression had a lower MACEs-free survival rate (long rank P<0.001) ( Figure 7 ). Multivariate Cox regression analysis was used to analyze the relationship between exosomal masc RNA and MACEs in ACS patients. After adjusting for age, diabetes, and LVEF, masc RNA was significantly associated with the occurrence of 1-year MACEs, with a HR of 2.959 and a 95% CI of 1.187-4.669 (P<0.001) (Table 6).

[0102] Table 6 Multivariate Cox regression model analysis of MACEs in ACS patients

[0103] HR 95% CI P-value HR 95% CI P* Age (years) 1.047 1.008-1.087 0.016 1.038 1.005-1.073 0.031 Gender (Male / Female) 1.577 0.702-3.543 0.270 hypertension 0.714 0.348-1.464 0.358 diabetes 0.873 0.357-2.136 0.766 0.529 0.198-1.412 0.204 Dyslipidemia (%) 1.024 0.479-2.188 0.951 LVEF (%) 0.954 0.929-0.980 0.001 0.955 0.931-0.979 <0.001 Number of brackets 0.995 0.564-1.754 0.986 Bracket length 1.008 0.986-1.030 0.494 MascRNA(log10) 3.357 2.042-5.520 <0.001 2.959 1.187-4.669 <0.001

[0104] *Adjusted for age, diabetes, and LVEF.

[0105] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of mascRNA in the preparation of any of the following products: (1) Diagnosis of acute coronary syndrome; (2) Screening for acute coronary syndrome; (3) Assess the prognosis of acute coronary syndrome.

2. The use according to claim 1, characterized in that: The expression level of mascRNA in patients with acute coronary syndrome was significantly higher than that in those with normal coronary arteries.

3. The use according to claim 1, characterized in that: The product includes at least one of a chip, a preparation, a kit, a test paper or a high-throughput sequencing platform.

4. The use according to claim 1, characterized in that: The product diagnoses, screens or evaluates the prognosis of acute coronary syndrome by detecting the expression level of mascRNA in a sample.

5. Use of a product for detecting mascRNA expression levels in the preparation of any of the following products: (1) Diagnosis of acute coronary syndrome; (2) Screening for acute coronary syndrome; (3) Assess the prognosis of acute coronary syndrome.

6. The use according to claim 5, characterized in that The products for detecting the expression level of mascRNA include: products for detecting the expression level of mascRNA through RT-PCR, fluorescent quantitative PCR, immunoassay, in situ hybridization, chip or high-throughput sequencing platform.

7. Application of mascRNA in the preparation of fluorescent quantitative PCR kit for diagnosing acute coronary syndrome.

8. The use according to claim 7, characterized in that: The kit comprises at least a pair of primers for specifically amplifying mascRNA, and the sequences are shown in SEQ ID NO.1 and SEQ ID NO.

2.

9. A fluorescent quantitative PCR kit for diagnosing acute coronary syndrome, characterized in that: The kit comprises at least a pair of primers for specifically amplifying mascRNA, and the sequences are shown in SEQ ID NO.1 and SEQ ID NO.

2.

10. The fluorescent quantitative PCR kit according to claim 9, characterized in that: The kit also includes a pair of primers for specifically amplifying the internal reference U6, the sequences of which are shown in SEQ ID NO.3 and SEQ ID NO.4.

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