Kit for predicting prognosis condition of cerebral hemorrhage patient

Abstract

The invention discloses a kit for predicting the prognostic condition of a cerebral hemorrhage patient, and relates to the field of biotechnology and medicine.The kit judges the prognostic condition of the cerebral hemorrhage patient by detecting the content of a biomarker anti-IL18bp in peripheral serum of the cerebral hemorrhage patient, and the prognostic condition of the patient can be predicted in advance. Therefore, the personalized treatment scheme of the patient is provided.

Description

Technical Field

[0001] The invention relates to the fields of biotechnology and medicine, and in particular to a kit for predicting the prognosis of patients with cerebral hemorrhage. Background Art

[0002] IL-18 is a member of the interleukin family and a pro-inflammatory cytokine that plays an important role in the inflammatory process. It can induce the production of a variety of inflammatory cytokines and chemokines, such as tumor necrosis factor-α (TNF-α), IL-1β, IL-6, etc., and then recruit and activate more inflammatory cells to the inflammatory site, amplifying the inflammatory response. IL-18 is also an immunomodulatory factor that can promote the proliferation and differentiation of T cells and enhance cellular immune responses.

[0003] Intracerebral hemorrhage (ICH) is an acute cerebrovascular disease with a high mortality and disability rate. Its prognosis is affected by multiple factors such as hematoma volume, location, secondary brain damage and systemic inflammatory response. At present, clinical prognosis evaluation mainly relies on imaging (such as CT / MRI) and neurological function scores (such as NIHSS, GCS), but these indicators are difficult to dynamically reflect the molecular pathological mechanism of brain tissue damage, especially in the regulation of inflammation. In addition, there are problems such as strong subjectivity in evaluation, high cost, and inability to accurately predict in the early stage. However, it is difficult to predict the prognosis in advance using long-term Barthel scores or mRS scores, resulting in delayed treatment plans. Therefore, a more objective, accurate and convenient prediction method that can reflect the changes in the disease earlier than imaging should be found, which has important clinical significance for the prognosis evaluation of patients with cerebral hemorrhage. Summary of the invention

[0004] In response to the above problems, the present invention provides an objective, accurate and convenient kit for predicting the prognosis of patients with cerebral hemorrhage, which can predict the prognosis of patients in advance to provide patients with personalized treatment plans.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: A kit for predicting the prognosis of patients with cerebral hemorrhage, wherein the biomarker detected by the kit is anti-IL18bp.

[0006] Preferably, the kit predicts the prognosis of patients with cerebral hemorrhage by detecting the content of the biomarker anti-IL18bp in the peripheral serum of patients with cerebral hemorrhage.

[0007] Furthermore, the prognosis prediction of the cerebral hemorrhage patient includes the prediction of daily living activities ability and the prediction of disability degree.

[0008] Furthermore, after the anti-IL18bp content is obtained by using the kit, the prognosis judgment equation is as follows: Y1=-0.0692X+111.0799; Y2=0.0023X+1.2279; Among them, X is the Anti-IL18bp content, Y1 is the score of daily living activities, and Y2 is the score of disability degree; Judgment rules: The lower the Y1 value, the lower the ability to perform daily activities and the worse the patient's prognosis; The higher the Y2 value, the higher the degree of disability and the worse the patient's prognosis.

[0009] Furthermore, the kit detects the content of anti-IL18bp by an indirect ELISA method.

[0010] Furthermore, it is characterized in that the kit is coated with IL-18bp antigen.

[0011] Furthermore, the kit also includes a freeze-dried standard, an HRP-antibody working solution, a TMB colorimetric substrate, a sample diluent, a reaction termination solution, a washing solution and a coating.

[0012] According to the composition of the test kit, the preparation method of the test kit is as follows: (1) After collecting peripheral blood from patients with cerebral hemorrhage, the supernatant is taken, and then diluted with a sample diluent to obtain a sample for use; (2) Wash the ELISA plate with 1X washing solution, add 100ul of sample or lyophilized standard, cover with film, and treat at 37℃ for 90min; (3) After washing, add 100ul 100X HRP-antibody working solution, cover with film, and treat at 37℃ for 30min; (4) After washing, add 90 μl of TMB colorimetric substrate, cover with film, and incubate at 37°C for 15 min. (5) Add 50ul of reaction stop solution and immediately read the OD value at 450nm. Calculate the formula based on the OD value of the standard to obtain the anti-IL18bp content of the sample to be tested.

[0013] Preferably, peripheral serum is collected from patients with cerebral hemorrhage on the 3rd to 14th day after onset of the disease.

[0014] Of course, the present invention also discloses the use of a reagent for detecting the expression level of anti-IL18bp in preparing a product for evaluating the prognosis of patients with cerebral hemorrhage, and the prognosis of cerebral hemorrhage is judged by detecting the expression level of anti-IL18bp in the patient's peripheral serum. Beneficial effects:

[0015] 1. The present invention evaluates the prognosis of patients with cerebral hemorrhage by detecting the relative expression of anti-IL18bp in the peripheral serum of patients, which has high clinical significance.

[0016] 2. The present invention provides a new method for the prognosis of cerebral hemorrhage, which can predict the patient's prognosis in advance, provide a reference basis for subsequent treatment plans and rehabilitation suggestions, and provide an opinion basis for formulating personalized treatments. It has broad application prospects in the field of medicine.

[0017] 3. The detection results of the present invention are reliable, the detection time is short, the detection method is simple and easy to operate, and the evaluation time is earlier than the evaluation table currently used in clinical practice, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 :Comparison of anti-IL18bp levels in peripheral serum of patients with cerebral hemorrhage and normal subjects; Figure 2 : Regression analysis of the anti-IL18bp content in the peripheral serum of patients with cerebral hemorrhage and the long-term Barthel score of the patients (P=0.02) (left); regression analysis of the anti-IL18bp content in the peripheral serum of patients with cerebral hemorrhage and the long-term mRS score of the patients (P=0.059) (right). DETAILED DESCRIPTION

[0019] The present invention will be described in detail below with reference to specific embodiments and accompanying drawings: Embodiment 1:

[0020] 1. Preparation of test kit Peripheral sera of 22 patients with cerebral hemorrhage randomly selected from the Department of Neurosurgery of the First Affiliated Hospital of Army Medical University on days 3-14 were collected between 2023 and 2024. Peripheral sera of 22 healthy people randomly selected from the Physical Examination Center of the First Affiliated Hospital of Army Medical University were collected as healthy samples.

[0021] (1) Centrifuge the peripheral serum sample at 2000g for 20 min, aspirate the supernatant and store it at -80℃, and dilute the serum sample 100 times with diluent for later use; (2) Equilibrate the ELISA plate at room temperature for 20 min and wash the plate twice with 1X washing solution; (3) Add 100 μl of sample or freeze-dried standard to each well, cover with film, and treat at 37°C for 90 min; (4) Wash the plate three times with 1X washing solution, add 100ul 100X HRP-antibody working solution to each well, cover with film, and treat at 37℃ for 30min; (5) Wash the plate 5 times with 1X washing buffer, add 90 μl TMB colorimetric substrate to each well, cover with film, and treat at 37°C for 15 min; (6) Add 50ul of reaction stop solution to each well and immediately read the OD value at 450nm. Calculate the formula based on the OD value of the standard to obtain the anti-IL18bp content of the sample to be tested.

[0022] The results obtained are Figure 1 As shown, the analysis results show that: The results showed that the level of anti-IL-18bp in the serum of patients with cerebral hemorrhage was significantly higher than that in the serum of healthy people, indicating that the level of anti-IL18bp in the peripheral serum of patients with cerebral hemorrhage can be used independently as a detection indicator to predict the prognosis of patients with cerebral hemorrhage.

[0023] 2. Prediction of prognosis of patients with cerebral hemorrhage: The prediction formula of Anti-IL18bp content and prognosis of patients with cerebral hemorrhage is as follows. Specifically, (1) Barthel prediction formula for daily living activities: Y1=-0.0692X1+111.0799; Among them, X1 is the Anti-IL18bp content, and Y1 is the score of daily living activities; Judgment rule: The lower the Y1 value, the lower the ability of daily living activities and the worse the patient's prognosis.

[0024] (2) Disability level (mRS) prediction formula: Y2=0.0023X2+1.2279; Among them, X2 is the Anti-IL18bp content, and Y2 is the disability degree score; Judgment rule: The higher the Y2 value, the higher the degree of disability and the worse the patient's prognosis.

[0025] 3. Verification of prediction results Peripheral serum samples of 4 patients with cerebral hemorrhage randomly selected from the Department of Neurosurgery, First Affiliated Hospital of Army Medical University, collected between July 2023 and February 2024, were collected on days 3 to 14. The Anti-IL18bp content X was detected according to the above method and kit, and the Y value was calculated according to formulas (1) and (2) to obtain the prediction results. Long-term telephone follow-up was conducted six months after the onset of the disease, and the patient self-care ability assessment scale (Barthel index) and modified Rankin scale score (mRS) were used for actual analysis. The results are shown in Table 2.

[0026] Calculation formula: (1) Barthel prediction formula: Y1=-0.0692X1+111.0799; Among them, X1 is the Anti-IL18bp content, and Y1 is the score of daily living activities; (2) Disability level (mRS) prediction formula: Y2=0.0023X2+1.2279; Among them, X2 is the Anti-IL18bp content, and Y2 is the disability degree score.

[0027] Table 2

[0028] Result analysis: According to the results in Table 2, the higher the anti-IL18bp content (X value) in the peripheral serum of patients with cerebral hemorrhage, the lower the long-term Barthel score (Y1) of the patients with cerebral hemorrhage (the lower the ability of daily living activities), and the higher the long-term mRS score (Y2) (the higher the degree of disability). For example, patient 1, Tang, had an anti-IL18bp content in peripheral serum of 1562.878 ng / ml, a Y1 value of 2.93, and a Y2 value of 4.82. It was judged that his prognosis was poor, his daily living ability was poor, and his degree of disability was high. This result was consistent with the results of the follow-up visit in October, indicating that the method and kit of the present invention can simply, conveniently and accurately judge the prognosis of patients with cerebral hemorrhage.

[0029] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. A kit for predicting the prognosis of a patient with cerebral hemorrhage, characterized in that: The biomarker detected by the kit is anti-IL18bp.

2. A kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 1, characterized in that: The kit determines the prognosis of patients with cerebral hemorrhage by detecting the content of the biomarker anti-IL18bp in the peripheral serum of patients with cerebral hemorrhage.

3. A kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 2, characterized in that: The prognosis prediction of the cerebral hemorrhage patient includes the prediction of daily living activity ability and the prediction of disability degree.

4. A kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 3, characterized in that: After the anti-IL18bp content is obtained using the kit, it is introduced into the prognosis judgment equation as follows: Y1=-0.0692X+111.0799; Y2=0.0023X+1.2279; Among them, X is the Anti-IL18bp content, Y1 is the score of daily living activities, and Y2 is the score of disability degree; Judgment rules: The lower the Y1 value, the lower the ability to perform daily activities and the worse the patient's prognosis; The higher the Y2 value, the higher the degree of disability and the worse the patient's prognosis.

5. A kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 4, characterized in that: The kit detects the content of anti-IL18bp by an indirect ELISA method.

6. A kit for predicting the prognosis of a patient with cerebral hemorrhage according to any one of claims 1 to 5, characterized in that: The kit is coated with IL-18bp antigen.

7. A kit for predicting the prognosis of a patient with cerebral hemorrhage as claimed in claim 6, characterized in that: The kit also includes a freeze-dried standard, an HRP-antibody working solution, a TMB color developing substrate, a sample diluent, a reaction termination solution, a washing solution and a film.

8. The method for preparing a kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 7, characterized in that: The method for making the detection kit is as follows: (1) After collecting peripheral blood from patients with cerebral hemorrhage, the supernatant is taken, and then diluted with a sample diluent to obtain a sample for use; (2) Wash the ELISA plate with 1X washing solution, add 100ul of sample or lyophilized standard, cover with film, and treat at 37℃ for 90min; (3) After washing, add 100ul 100X HRP-antibody working solution, cover with film, and treat at 37℃ for 30min; (4) After washing, add 90 μl of TMB colorimetric substrate, cover with film, and incubate at 37°C for 15 min. (5) Add 50ul of reaction stop solution and immediately read the OD value at 450nm. Calculate the formula based on the OD value of the standard to obtain the anti-IL18bp content of the sample to be tested.

9. The method for preparing a kit for predicting the prognosis of a patient with cerebral hemorrhage according to claim 8, characterized in that: Peripheral serum was collected from patients with cerebral hemorrhage on the 3rd to 14th day after onset.

10. Use of a reagent for detecting the expression level of anti-IL18bp in the preparation of a product for the prognosis assessment of patients with cerebral hemorrhage, characterized in that: The prognosis of cerebral hemorrhage is determined by detecting the expression level of anti-IL18bp in the patient's peripheral serum.

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