Use of a product for detecting biomarkers in preparing a product for diagnosing acute myocardial infarction or its complications

Abstract

The present invention relates to the field of biomedicine technology, and in particular to the use of a product for detecting biomarkers in the preparation of a product for the diagnosis of acute myocardial infarction or its complications. The present invention provides a product for detecting biomarkers for the preparation of a product for auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of acute myocardial infarction or its complications, wherein the biomarker includes Spondin-1 protein. The present invention has found that Spondin-1 protein has high specificity and sensitivity as a biomarker for the diagnosis of acute myocardial infarction, and has high accuracy in diagnosing acute myocardial infarction. It can be used as a target for auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of acute myocardial infarction or its complications. Therefore, the product for detecting Spondin-1 protein can be used to prepare a product for auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of acute myocardial infarction or its complications.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to use of a product for detecting biomarkers in preparing a product for diagnosing acute myocardial infarction or its complications. Background Art

[0002] Cardiovascular disease carries a high incidence and mortality rate across the human population. Currently, these rates continue to rise, making it the leading threat to human health. Acute myocardial infarction (AMI) is a leading cause of death and the leading cause of death among cardiovascular diseases. Therefore, improving the prevention, diagnosis, and treatment of AMI is crucial. Summary of the Invention

[0003] Therefore, the technical problem to be solved by the present invention is to provide a product for detecting biomarkers for use in preparing a product for diagnosing AMI or its complications.

[0004] To this end, the present invention provides the following technical solutions:

[0005] A product for detecting biomarkers is used in the preparation of a product for auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of AMI or its complications, wherein the biomarker includes Spondin-1 protein.

[0006] Optionally, the invention also includes use in preparing products for auxiliary diagnosis or diagnosis of AMI prognosis.

[0007] Optionally, the method also includes use in preparing a product for predicting myocardial infarction area.

[0008] Optionally, the complications include AMI combined with cardiac rupture and AMI combined with heart failure.

[0009] Optionally, the product for preparing auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of AMI or its complications includes a kit, a test paper, a reagent or an instrument.

[0010] Optionally, the product for detecting biomarkers includes a kit, a test paper, a reagent or an instrument.

[0011] Optionally, the sample to be tested for detecting biomarkers includes plasma.

[0012] Optionally, the sample to be tested is peripheral blood plasma.

[0013] The technical solution of the present invention has the following advantages:

[0014] 1. The present invention provides a product for detecting biomarkers for use in preparing products for auxiliary diagnosis, diagnosis, risk assessment, or prediction of clinical prognosis of AMI or its complications, wherein the biomarker includes Spondin-1 protein. The present invention's study found that Spondin-1 protein was highly expressed in AMI, significantly higher than the expression level of Spondin-1 protein in the control group, indicating that the expression level of Spondin-1 protein was significantly correlated with AMI. At the same time, combined with ROC curve analysis, it was found that the AUC value of Spondin-1 protein for AMI diagnosis was 0.867. As a biomarker for AMI diagnosis, Spondin-1 protein has high specificity and sensitivity, and has high accuracy in diagnosing AMI. It can be used as a target for auxiliary diagnosis, diagnosis, risk assessment, or prediction of clinical prognosis of AMI. Further studies have found that plasma Spondin-1 levels are correlated with myocardial infarction size, with elevations more pronounced in patients with an infarction area greater than 30%. AMI patients with heart failure have higher plasma Spondin-1 levels compared to those without heart failure, and AMI patients with myocardial rupture have higher plasma Spondin-1 levels compared to those without. These findings suggest that Spondin-1 can be used as a biomarker to diagnose AMI and predict AMI complications (such as heart failure and myocardial rupture). Therefore, products for detecting Spondin-1 protein can be used to develop products for auxiliary diagnosis, diagnosis, risk assessment, or clinical prognosis prediction of AMI or its complications. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1 is the standard curve of Spondin-1 in the embodiment of the present invention;

[0017] Figure 2 The results of Spondin-1 protein concentration detection in the experimental group and the control group in the embodiment of the present invention are shown in the figure. In the figure, Control represents the control group, * represents P < 0.05, *** represents P < 0.001, and **** represents P < 0.0001.

[0018] Figure 3 is the ROC curve analysis result in the embodiment of the present invention;

[0019] Figure 4This is the correlation result between the myocardial infarction area and the Spondin-1 protein (SPON1 in the figure) concentration of the sample to be tested in the experimental group of the embodiment of the present invention;

[0020] Figure 5 This is the comparison result of Spondin-1 protein concentration between AMI patients with heart failure and AMI patients without heart failure in the experimental group of the embodiment of the present invention; * indicates P < 0.05, *** indicates P < 0.001, and **** indicates P < 0.0001;

[0021] Figure 6 This is the comparison result of Spondin-1 protein concentration between patients with AMI complicated with cardiac rupture and patients with AMI without cardiac rupture in the control group and the experimental group in the embodiment of the present invention; * indicates P < 0.05, *** indicates P < 0.001, and **** indicates P < 0.0001. DETAILED DESCRIPTION

[0022] The following examples are provided for a better understanding of the present invention and are not intended to limit the best mode of implementation. They do not limit the content and scope of protection of the present invention. Any product identical or similar to the present invention obtained by anyone under the guidance of the present invention or by combining the features of the present invention with other prior arts shall fall within the scope of protection of the present invention.

[0023] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in this field can be used. If the manufacturer of the reagents or instruments is not specified, they are all commercially available conventional reagents.

[0024] Example

[0025] 1. Experimental subjects

[0026] In this study, the diagnostic and predictive value of Spondin-1 in AMI and its complications was tested in three populations: (1) Population 1: This population included 30 AMI patients and 60 unstable angina (UA) patients. The diagnostic criteria for AMI are: elevated myocardial injury markers and at least one of the following evidence of myocardial ischemia: ① significant clinical manifestations of myocardial ischemia; ② pathological Q waves on the electrocardiogram; ③ ST segment elevation on the electrocardiogram. The diagnostic criteria for UA are consistent with the diagnostic criteria for unstable angina published by the European Society of Cardiology in 2020. Among them, all 30 AMI patients underwent cardiac magnetic resonance imaging, and the magnetic resonance imaging data were analyzed by physicians. The infarct size was defined as the high signal area in the delayed enhancement image, whose myocardial grayscale threshold exceeded 5 standard deviations of the normal myocardium distal to the infarct site. (2) Population 2: This population included 22 patients with AMI complicated by cardiac rupture, 40 patients with AMI without cardiac rupture, and 70 patients with stable coronary artery disease (SCAD). The diagnostic criteria for patients with cardiac rupture (at least one of the following evidences): ① cardiac rupture confirmed by autopsy or surgery; ② cardiac rupture confirmed by echocardiography, right heart catheterization, and ventriculography; ③ cardiac arrest caused by electromechanical dissociation without a history of heart failure, and pericardial hematoma confirmed by echocardiography; ④ persistent hypotension, pericardial effusion confirmed by echocardiography (pericardial effusion ≥10 mm with right atrial compression) and bloody pericardial effusion on pericardiocentesis. The diagnostic criteria for SCAD were in accordance with the diagnostic criteria for stable coronary artery disease published by the European Society of Cardiology in 2013. (3) Population 3: This population included 24 patients with AMI complicated by heart failure and 133 patients with AMI without heart failure. Diagnostic criteria for heart failure: Refer to the classification and diagnostic criteria for heart failure in the 2018 Chinese Guidelines for the Diagnosis and Treatment of Heart Failure. All of the above cases were treated at Beijing Anzhen Hospital, affiliated with Capital Medical University.

[0027] 2. Experimental methods

[0028] (1) Preparation of test samples: Peripheral blood was collected from the experimental group and the control group and placed in EDTA anticoagulant tubes. Plasma was separated by centrifugation within 30 minutes after collection and stored in a -80°C refrigerator for future use.

[0029] (2) Detect the sample in step (1) according to the steps of the ELISA kit (Human Spondin-1 (SPON1) ELISA kit (CSB-EL022599HU, Wuhan Huamei Biotechnology). The operation process is as follows:

[0030] 1) After equilibration at room temperature for 60 minutes, remove the desired strips from the aluminum foil bag. Seal the remaining strips in a ziplock bag and return them to 4°C.

[0031] 2) Set up standard wells, blank wells, and sample wells, and add 50 μL of standard of different concentrations to each standard well.

[0032] 3) Add 50 μL of the sample to be tested to the sample well and 50 μL of the sample diluent to the blank well.

[0033] 4) Add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each of the blank wells, standard wells, and sample wells, seal the reaction wells with a sealing film, and incubate at 37°C in a constant temperature box for 60 min.

[0034] 5) Discard the liquid, pat dry with absorbent paper, fill each well with washing solution (350 μL), let it stand for 1 minute, shake off the washing solution, pat dry with absorbent paper, and repeat 5 times.

[0035] 6) Add 50 μL of substrate A and substrate B to each well and incubate at 37°C in the dark for 15 min.

[0036] 7) Add 50 μL of stop solution to each well and measure the OD value of each well at a wavelength of 450 nm within 15 minutes.

[0037] A standard curve was drawn based on the OD values ​​of the blank wells and the standard wells. The Spondin-1 protein concentration in the test sample was calculated using the standard curve. Each sample in the experimental and control groups was repeated three times.

[0038] 3. Statistical Analysis Methods

[0039] Data were analyzed using SPSS 26.0 statistical software. Intergroup comparisons were performed using independent t-tests and rank-sum tests. Normally distributed data were expressed as mean ± standard deviation, while those not normally distributed were expressed as median and interquartile ranges. Receiver operating characteristic (ROC) statistics were used for diagnostic testing.

[0040] 4. Results

[0041] 4.1 Experimental results of AMI

[0042] 4.1.1 Correlation between AMI and Spondin-1 Protein

[0043] The standard curve of Spondin-1 is as follows Figure 1 As shown in FIG, the Spondin-1 protein concentrations in the test samples of the AMI group (30 AMI patients) and the control group (60 UA patients) obtained by applying the standard curve are shown in FIG. Figure 2As shown in Figure 2, the expression of Spondin-1 protein in AMI patients was 1.93 times that in the control group (P<0.001), indicating that Spondin-1 was highly expressed in AMI. The ROC analysis results of Spondin-1 protein for AMI diagnosis are shown in Figure 2. Figure 3 The results show an AUC of 0.867 [0.795-0.938], p < 0.001, and a sensitivity of 0.906. These results demonstrate that spondin-1 protein has high sensitivity as a biomarker for the diagnosis of AMI and can serve as a target for auxiliary diagnosis, diagnosis, risk assessment, or clinical prognosis prediction of AMI. Therefore, products detecting spondin-1 protein can be used to develop products for auxiliary diagnosis, diagnosis, risk assessment, or clinical prognosis prediction of AMI.

[0044] 4.1.2 Correlation between Myocardial Infarction Size and Spondin-1 Protein

[0045] The correlation analysis was performed between the Spondin-1 protein concentration in the samples of the AMI group (30 AMI patients) and the myocardial infarction area (%) of the samples. Figure 4 As shown in the data, plasma Spondin-1 is correlated with myocardial infarction size, and the increase is more significant in patients with myocardial infarction area >30%, indicating that Spondin-1 can be used as a biomarker to predict myocardial infarction area in acute myocardial infarction.

[0046] 4.2 Experimental results of heart failure

[0047] The plasma Spondin-1 levels in AMI patients with heart failure (24 AMI patients with heart failure, experimental group) and AMI patients without heart failure (133 AMI patients without heart failure, control group) were compared. Figure 5 As shown in the data, the plasma Spondin-1 level is higher in AMI patients with concomitant heart failure, indicating that Spondin-1 can be used as a biomarker for heart failure complicated by acute myocardial infarction. Products for detecting Spondin-1 protein can be used to prepare products for auxiliary diagnosis, diagnosis, risk assessment or prediction of clinical prognosis of heart failure complicated by acute myocardial infarction.

[0048] 4.3 Experimental results of heart rupture

[0049] The plasma levels of Spondin-1 were compared in a control group (70 patients with stable coronary artery disease (SCAD)), patients with AMI complicated by cardiac rupture (22 patients with AMI complicated by cardiac rupture), and patients with AMI without cardiac rupture (40 patients with AMI without cardiac rupture). Figure 6As shown, patients with AMI complicated by cardiac rupture have higher plasma spondin-1 levels. The above shows that spondin-1 can be used as a biomarker for cardiac rupture, a complication of acute myocardial infarction. Products that detect spondin-1 protein can be used to prepare products for auxiliary diagnosis, diagnosis, risk assessment, or clinical prognosis prediction of cardiac rupture, a complication of acute myocardial infarction.

[0050] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will readily appreciate that other variations or modifications based on the above descriptions are possible. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. Use of a product for detecting biomarkers in the preparation of a product for auxiliary diagnosis, diagnosis or risk assessment of acute myocardial infarction, characterized in that: The biomarkers include Spondin-1 protein.

2. The use according to claim 1, characterized in that It also includes use in preparing products for predicting myocardial infarction area.

3. The use according to claim 1 or 2, characterized in that The products for preparing auxiliary diagnosis, diagnosis or risk assessment of acute myocardial infarction include kits, test strips, reagents or instruments.

4. The use according to claim 1 or 2, characterized in that The products for detecting biomarkers include kits, test strips, reagents or instruments.

5. The use according to claim 1 or 2, characterized in that The sample to be tested for detecting the biomarker includes plasma.

6. The use according to claim 5, characterized in that The sample to be tested is peripheral blood plasma.

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