Application of HFREP1 protein in preparing a kit for diagnosing bronchial asthma

Abstract

The present invention discloses an application of HFREP1 protein in preparing a kit for diagnosing bronchial asthma. The present invention finds that there is a significant difference in the HFREP1 protein content in the serum of healthy subjects and bronchial asthma patients, and the HFREP1 protein content in the serum of bronchial asthma patients is significantly higher than that of healthy subjects. Further diagnostic efficacy tests show that HFREP1 protein can effectively diagnose and distinguish healthy subjects from bronchial asthma patients with high diagnostic accuracy, and can be used to develop a kit for diagnosing bronchial asthma, which is significantly more objectivity than existing diagnostic methods.

Description

Technical Field

[0001] The invention belongs to the field of disease diagnosis, relates to the discovery and application of disease diagnosis markers, and specifically relates to the application of HFREP1 protein in the preparation of a kit for diagnosing bronchial asthma. Background Art

[0002] Bronchial asthma is a chronic inflammatory disease of the airways involving multiple cells and cell components. If bronchial asthma is not diagnosed and treated in time, irreversible airway narrowing and airway remodeling may occur as the disease progresses. When asthma is controlled, most patients rarely experience asthma attacks, and severe asthma attacks are even rarer. Therefore, reasonable prevention and treatment are crucial. The prevalence of asthma in adults in my country is 1.24%, and the prevalence of adult men and women is roughly the same. About 40% of patients have a family history.

[0003] In order to improve the objectivity of bronchial asthma diagnosis, the present invention is proposed. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an application of HFREP1 protein in the preparation of a kit for diagnosing bronchial asthma.

[0005] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0006] A use of HFREP1 protein in preparing a kit for diagnosing bronchial asthma.

[0007] Preferably, the kit is a serum detection kit.

[0008] Preferably, the kit contains a detection reagent for HFREP1 protein.

[0009] Beneficial effects:

[0010] The present invention found that there was a significant difference in the HFREP1 protein content in the serum of healthy subjects and bronchial asthma patients, and the HFREP1 protein content in the serum of bronchial asthma patients was significantly higher than that of healthy subjects. Further diagnostic efficacy tests showed that HFREP1 protein can effectively diagnose and distinguish healthy subjects from bronchial asthma patients with high diagnostic accuracy, and can be used to develop a kit for diagnosing bronchial asthma, which is significantly more objective than existing diagnostic methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 ROC curve for distinguishing healthy subjects from bronchial asthma patients in the training set for HFREP1 protein diagnosis;

[0012] Figure 2This is the sample distribution diagram for predicting healthy subjects vs bronchial asthma patients based on HFREP1 protein in the validation set. DETAILED DESCRIPTION

[0013] The essential contents of the present invention are described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0014] 1. Experimental samples and reagents

[0015] 34 healthy subjects and 29 patients with bronchial asthma were collected. Healthy subjects were normal people with healthy physical examinations, and patients with bronchial asthma were diagnosed according to the diagnostic criteria of the 2020 Bronchial Asthma Prevention and Treatment Guidelines. The two groups were matched in age, gender, and body mass index, with no significant differences. The subjects or patients were randomly divided into training set samples (18 healthy subjects and 18 patients with bronchial asthma) and validation set samples (16 healthy subjects and 11 patients with bronchial asthma).

[0016] Exclusion criteria:

[0017] ① Patients with other acute diseases or uncontrolled chronic diseases; ② Patients with serious primary diseases such as cardiovascular, liver, kidney and hematopoietic system; ③ Patients with mental illness and unable to cooperate; ④ Pregnant or lactating women, or those preparing for pregnancy; ⑤ Those who have participated in other clinical trials within the past month; ⑥ Those who are unwilling to accept this study.

[0018] Main experimental reagents:

[0019] The ELISA reagent for measuring HFREP1 protein was purchased from RayBiotech., Inc.

[0020] 2. Experimental Methods

[0021] 1. Collection and storage of serum samples

[0022] Fasting peripheral blood was collected from healthy subjects and bronchial asthma patients in the early morning and placed in a test tube without anticoagulant. The blood was naturally coagulated for 30-60 minutes at room temperature. After the blood coagulated, it was centrifuged at 2000 rpm for 10 minutes. The upper clear serum liquid was carefully aspirated into a sterile lyophilized tube, labeled and stored in a -80℃ refrigerator for later use.

[0023] 2. ELISA determination of HFREP1 protein content in serum

[0024] The content of HFREP1 protein in each serum sample was determined strictly according to the ELISA reagent assay instructions.

[0025] 3. Data processing method

[0026] The ROC curve of HFREP1 protein in the test set for distinguishing healthy subjects from patients with bronchial asthma was established, and the area under the curve (AUC) and 95% confidence interval were calculated. Logistic regression was used to establish a regression equation to generate a set of new variables logit[P]. After logit[P]=Ln [p / (1-p)] The predicted probability p was obtained by transformation, and the ROC curve analysis was performed on the new variable. In the validation set, the best cut-off value obtained by the ROC curve was used as the threshold to calculate the diagnostic accuracy of HFREP1 for bronchial asthma.

[0027] 3. Experimental Results

[0028] 1. Differences in serum levels of HFREP1 protein between healthy subjects and patients with bronchial asthma

[0029] In the training set, there were significant differences in the levels of HFREP1 protein in the serum of healthy subjects VS patients with bronchial asthma. The absolute content of HFREP1 in the serum of patients with bronchial asthma was significantly increased. The measurement results are shown in Table 1.

[0030] Table 1

[0031] Group HFREP1 content (ng / mL) Bronchial asthma 542.88±515.33 Healthy subjects 29.53±16.39

[0032] 2. ROC curve of HFREP1 protein diagnosis for distinguishing healthy subjects from bronchial asthma patients

[0033] The HFREP1 protein content (X) in the training set samples was used as the independent variable, and the groups (i.e., healthy subjects and bronchial asthma patients) were used as the dependent variable. Logistic regression was performed on the content of HFREP1 protein in serum samples of bronchial asthma and healthy subjects, and the logistic regression equation Ln was obtained. [p / (1-p)] =0.075X-4.607. Then, the absolute content of HFREP1 protein in each serum sample is substituted into the logistic regression equation to obtain the regression prediction probability p of each serum sample. The possible regression prediction probability p is used as the diagnostic point to calculate the sensitivity and specificity, and the ROC curve is drawn accordingly (e.g. Figure 1 The AUC is 0.9568, the sensitivity is 1, and the specificity is 0.9444. The Wieden index is calculated according to the coordinates of the ROC curve = specificity + sensitivity - 1. The predicted probability p value corresponding to the maximum value of the Wieden index is the best cut-off value of 0.4556 for distinguishing healthy subjects from bronchial asthma patients.

[0034] The area under the ROC curve (AUC) has been widely recognized as an inherent accuracy indicator for evaluating the authenticity of diagnostic tests. When AUC is 0.5, it has no diagnostic significance; when AUC is between 0.5 and 0.7, it indicates a low diagnostic accuracy; when AUC is between 0.7 and 0.9, it indicates a medium diagnostic accuracy; when AUC>0.9, it indicates a high diagnostic accuracy.

[0035] 3. Verify the accuracy of HFREP1 protein in diagnosing bronchial asthma

[0036] In the validation set, the best cut-off value obtained above was used as the diagnostic threshold to predict the serum samples of bronchial asthma patients VS healthy subjects. The number of correctly predicted samples divided by the total number of samples was the accuracy of HFREP1 protein in distinguishing bronchial asthma patients VS healthy subjects. The accuracy of HFREP1 protein in distinguishing bronchial asthma patients VS healthy subjects was 74.07% (20 / 27). The sample distribution diagram is shown in Figure 2 shown.

[0037] In summary, there is a significant difference in the HFREP1 protein content in the serum of healthy subjects and bronchial asthma patients, and the HFREP1 protein content in the serum of bronchial asthma patients is significantly higher than that of healthy subjects. Further diagnostic efficacy tests show that HFREP1 protein can effectively diagnose and distinguish healthy subjects from bronchial asthma patients with high diagnostic accuracy, and can be used to develop a kit for diagnosing bronchial asthma, which is significantly more objective than existing diagnostic methods.

[0038] The purpose of the above-mentioned embodiments is to specifically introduce the essential content of the present invention, but those skilled in the art should know that the protection scope of the present invention should not be limited to the specific embodiments.

Claims

1. Use of HFREP1 protein in preparing a kit for diagnosing bronchial asthma.

2. The use according to claim 1, wherein the kit is a serum detection kit.

3. The use according to claim 1, wherein the kit contains a detection reagent for HFREP1 protein.

Images