Use of cfhr5 protein in the preparation of a kit for diagnosing the differentiation between prostate cancer and benign prostate hyperplasia

Abstract

The application discloses application of CFHR5 protein in preparation of a kit for diagnosing and distinguishing prostate cancer and benign prostatic hyperplasia. It is found by the application that the CFHR5 protein has high diagnostic value for distinguishing prostate cancer and benign prostatic hyperplasia, the accuracy of diagnosis and distinction is high, and the CFHR5 protein has the prospect of being developed into a kit for diagnosing and distinguishing prostate cancer and benign prostatic hyperplasia.

Description

Technical Field

[0001] This invention belongs to the field of biochemistry and relates to the application of biomarkers in disease diagnosis, specifically the application of CFHR5 protein in the preparation of a diagnostic kit to differentiate between prostate cancer and benign prostatic hyperplasia. Background Technology

[0002] Prostate cancer (PCa) is the most common malignant tumor in men. In recent years, with population aging and changes in lifestyle, the incidence of prostate cancer in my country has shown a continuous upward trend. Because early symptoms of prostate cancer are not obvious, most prostate cancer patients in my country are diagnosed at a stage where local progression or distant metastasis has already occurred, leading to a poor prognosis. Although prostate-specific antigen (PSA) has been widely used for the early diagnosis of prostate cancer, elevated plasma PSA levels can also be seen in non-tumor states such as benign prostatic hyperplasia (BPH). Due to its poor specificity, PSA often leads to overdiagnosis, failing to meet clinical needs. Therefore, finding other ideal biomarkers to combine with PSA for joint diagnosis, and improving the ability of PSA to differentiate between prostate cancer and benign prostatic hyperplasia, is key to promoting accurate prostate cancer screening.

[0003] Plasma is rich in a variety of proteins that participate extensively in multiple biological processes, including immunity, coagulation, growth signaling, and nutrition. Many diseases can lead to changes in the quantity and structure of plasma proteins, which are of great significance for disease diagnosis. In recent years, with the rapid development of proteomics, several plasma proteins have been revealed as potential biomarkers for tumor diagnosis, becoming an important approach to finding diagnostic biomarkers for diseases. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide the application of CFHR5 protein in the preparation of a diagnostic kit for differentiating prostate cancer from benign prostatic hyperplasia.

[0005] The above-mentioned objective of this invention is achieved through the following technical solution:

[0006] Application of CFHR5 protein in the preparation of diagnostic kits that differentiate between prostate cancer and benign prostatic hyperplasia.

[0007] A diagnostic kit for differentiating prostate cancer from benign prostatic hyperplasia, containing a reagent for detecting the CFHR5 protein.

[0008] Beneficial effects:

[0009] This invention has found that the CFHR5 protein has high diagnostic value in differentiating between prostate cancer and benign prostatic hyperplasia, with a high accuracy rate in the diagnosis, and shows promise for development into a diagnostic kit for differentiating between prostate cancer and benign prostatic hyperplasia. Attached Figure Description

[0010] Figure 1 To test the difference in plasma CFHR5 protein expression between patients with prostate cancer and benign prostatic hyperplasia;

[0011] Figure 2 To train the ROC curve analysis and diagnostic threshold (cutoff) for diagnosing and differentiating prostate cancer from benign prostatic hyperplasia patients using concentrated plasma CFHR5 protein. Detailed Implementation

[0012] The following describes the substantive content of the present invention in detail with reference to embodiments, but this does not limit the scope of protection of the present invention.

[0013] Example 1: Diagnostic efficacy of plasma CFHR5 protein in differentiating between prostate cancer and benign prostatic hyperplasia

[0014] I. Experimental Samples

[0015] Thirty-nine patients with prostate cancer and 39 patients with benign prostatic hyperplasia (BPH) from Changzhou First People's Hospital were randomly divided into a test set, a training set, and a validation set. The test set consisted of 15 patients with prostate cancer and 15 patients with BPH; the training set consisted of 12 patients with prostate cancer and 12 patients with BPH; and the validation set consisted of 12 patients with prostate cancer and 12 patients with BPH.

[0016] Inclusion criteria: Prostate cancer patients were those diagnosed by pathology and who had not received radiotherapy or chemotherapy; Benign prostatic hyperplasia (BPH) patients were those whose BPH lesions were confirmed as benign by biopsy.

[0017] Exclusion criteria: ① Not meeting the relevant diagnostic criteria for prostate cancer and benign prostatic hyperplasia; ② Having other prostate diseases, such as prostatitis; ③ Having other serious liver, kidney, heart, lung, and hematopoietic system diseases.

[0018] II. Experimental Methods

[0019] 1. Collection and storage of plasma samples

[0020] Five mL of peripheral venous blood was collected from the subjects using vacuum anticoagulation (EDTA) blood collection tubes. After standing for 2 hours, the blood cells and plasma were separated by centrifugation at 4,000 rpm for 20 minutes. The blood was then transferred to 2 mL Eppendorf tubes, and the plasma was stored at -80°C for long-term storage.

[0021] 2. Determination of CFHR5 protein content

[0022] The levels of CFHR5 protein in the plasma of patients with prostate cancer and benign prostatic hyperplasia were detected using enzyme-linked immunosorbent assay (ELISA). The ELISA kit was purchased from Shanghai Enzyme-linked Biotechnology (No. ml605809). The plasma CFHR5 levels were determined according to the kit's instructions. The specific experimental steps are as follows: Add the standard or sample to a 96-well microplate and incubate at 37°C for 1 hour. CFHR5 in the plate binds to the CFHR5 antibody coated in the microplate. Discard the liquid, add biotinylated CFHR5 antibody, and incubate at 37°C for 1 hour. Discard the liquid and wash three times with washing buffer to remove unbound biotinylated antibody. Add HRP (peroxidase)-labeled avidin and incubate at 37°C for 30 minutes to allow avidin to completely bind to biotin. Discard the liquid and wash five times with washing buffer. Add TMB substrate and incubate at 37°C in the dark for 10-20 minutes. TMB turns blue under the catalysis of HRP. Measure the absorbance at 450 nm. The absorbance is directly proportional to the CFHR5 protein content.

[0023] 3. Data processing methods

[0024] The expression levels of the target protein were expressed as mean ± standard deviation, and the Student's t-test was used to compare the differences in protein levels between different groups. The diagnostic value of CFHR5 protein for prostate cancer was evaluated by plotting receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC) and its 95% confidence interval (CI). An AUC of 0.5 indicated no diagnostic significance; an AUC of 0.5–0.7 indicated low diagnostic accuracy; an AUC of 0.7–0.9 indicated moderate diagnostic accuracy; and an AUC greater than 0.9 indicated high diagnostic accuracy. The DeLong's test was used to compare the differences in AUC among different models to comprehensively evaluate the application value of plasma CFHR5 as a diagnostic biomarker for prostate cancer.

[0025] The diagnostic evaluation indicators are calculated as follows:

[0026]

[0027] Sensitivity, also known as the true positive rate, is the probability that a person actually has the disease and is correctly diagnosed as having the disease by a diagnostic test. Sensitivity = [a / (a+c)] × 100%.

[0028] Specificity, also known as the true negative rate, is the probability that a diagnostic test correctly identifies a person as disease-free when they are actually disease-free. Specificity = [d / (b+d)] × 100%.

[0029] III. Experimental Results

[0030] 1. Differences in CFHR5 protein levels among patients with different diseases

[0031] In the test set, the plasma CFHR5 protein level was significantly upregulated in prostate cancer patients compared to patients with benign prostatic hyperplasia. The results are shown in the table below. Figure 1 As shown.

[0032]

[0033] 2. ROC curve analysis of CFHR5 protein in differentiating prostate cancer from benign prostatic hyperplasia

[0034] In the training set, the plasma CFHR5 protein levels of each patient were measured, ROC curves were plotted, and the area under the curve (AUC) and its 95% CI were calculated to evaluate the diagnostic value of CFHR5 protein for prostate cancer. ROC analysis results showed that in the training set, the AUC value of plasma CFHR5 in differentiating prostate cancer from benign prostatic hyperplasia patients was as high as 0.847 (sensitivity 0.750, specificity 0.833), with an optimal cutoff value of 2681.20. (See [link to relevant documentation]). Figure 2 .

[0035] 3. Verify the accuracy of CFHR5 protein alone in diagnosing and differentiating between prostate cancer and benign prostatic hyperplasia.

[0036] In the validation set, the CFHR5 protein level in the plasma of each patient was measured. The cutoff value of 2681.20 obtained from the training set was used as the diagnostic threshold. Patients with a cutoff below this threshold were predicted to have benign prostatic hyperplasia (BPH), while those above were predicted to have prostate cancer. The accuracy of CFHR5 protein in differentiating between prostate cancer and BPH was calculated by dividing the number of correctly predicted samples by the total number of samples. The accuracy of CFHR5 protein in differentiating between prostate cancer and BPH was 83.33% (20 / 24).

[0037] The above experimental results indicate that CFHR5 protein has high diagnostic value in differentiating between prostate cancer and benign prostatic hyperplasia, with a high accuracy rate in diagnosis and differentiation, and shows promise for development into a diagnostic kit for differentiating between prostate cancer and benign prostatic hyperplasia.

[0038] Example 2: Diagnostic kit for differentiating between prostate cancer and benign prostatic hyperplasia

[0039] A diagnostic kit for differentiating prostate cancer from benign prostatic hyperplasia, containing a reagent for detecting the CFHR5 protein.

[0040] The purpose of the above embodiments is to specifically illustrate the substantive content of the present invention, but those skilled in the art should know that the scope of protection of the present invention should not be limited to the specific embodiments.

Claims

1. Application of CFHR5 protein detection reagent in the preparation of a diagnostic kit to differentiate between prostate cancer and benign prostatic hyperplasia.

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