Biomarker for systemic sclerosis, and detection method therefor and use thereof

Abstract

A biomarker for systemic sclerosis, and a detection method therefor and the use thereof. The biomarker is an anti-PRMT5 antibody. The use of PRMT5 in the preparation of a drug or agent for detecting or treating systemic sclerosis, wherein patients with systemic sclerosis are distinguished from healthy controls and patients with other autoimmune diseases by means of detecting the anti-PRMT5 antibody level. The anti-PRMT5 antibody levels in patients with systemic sclerosis are closely correlated with the outcome of skin and pulmonary fibrosis in the patients, and the biomarker has potential value in prognosis evaluation. The detection method using enzyme-linked immunosorbent assay is simple, easy to perform, convenient for translation, and suitable for wide promotion in primary hospitals.

Description

A systemic sclerosis biomarker and its detection method and application Technical Field

[0001] The present invention belongs to the field of medical testing and biological technology, and relates to a systemic sclerosis biomarker and a detection method and application thereof, and specifically to a novel systemic sclerosis biomarker anti-PRMT5 antibody and a detection method and application thereof. Background Art

[0002] Systemic sclerosis (SSc) is an autoimmune disease that can affect multiple organs, including the skin, lungs, heart, digestive tract, and kidneys. The global prevalence of SSc is 17.6 per 100,000 people, with an overall incidence of 1.4 per 100,000 person-years.

[0003] Clinical practice demonstrates the complex and diverse manifestations of systemic sclerosis, including skin swelling and hardening, musculoskeletal pain, pulmonary fibrosis, digestive system symptoms, and renal crisis. Because disease-related organ damage is often irreversible, early identification, prognosis, and timely intervention are crucial. However, the diagnosis and assessment of SSc remain challenging in clinical practice, with a lack of objective assessment methods and models. Therefore, in-depth research into the pathogenesis of SSc and the discovery of novel biomarkers are crucial for the diagnosis and treatment of this disease.

[0004] Autoantibodies are valuable biomarkers for the diagnosis and treatment of systemic sclerosis (SSc). For example, anti-topoisomerase I antibodies (ATA) are associated with pulmonary fibrosis and poor prognosis, while anti-RNA polymerase antibodies (ARA) are more common in patients with diffuse SSc and are associated with renal crisis. However, clinical studies have shown that because SSc is a highly heterogeneous disease, up to 17% of SSc patients are negative for known specific antibodies. Furthermore, whether these autoantibodies play a pathogenic role in SSc remains to be determined.

[0005] Based on the current status of the prior art, the inventors of this application intend to provide a systemic sclerosis biomarker and its detection method and application, especially a novel systemic sclerosis biomarker anti-PRMT5 antibody and its detection method and application.

[0006] Summary of the Invention

[0007] The present invention aims to solve the technical problems existing in the prior art and provide a novel biomarker for systemic sclerosis and its applications. In particular, it provides a novel biomarker for systemic sclerosis, an anti-PRMT5 antibody, a detection method thereof, and its applications.

[0008] The present invention, in one aspect, first discovered that anti-PRMT5 antibodies are elevated in the serum of SSc patients and can serve as a diagnostic marker for SSc. The present invention further provides a method for detecting anti-PRMT5 antibodies in systemic sclerosis, comprising detecting the level of anti-PRMT5 antibodies in samples from systemic sclerosis patients using an enzyme-linked immunosorbent assay (ELISA).

[0009] In a second aspect of the present invention, research has found that anti-PRMT5 antibody levels are correlated with the progression of mRSS (metastatic RSS) and high-resolution CT scores on the lungs in patients, demonstrating potential prognostic value. The present invention also provides a method for detecting anti-PRMT5 antibodies in the peripheral blood of patients with systemic sclerosis, including the use of anti-PRMT5 antibodies in diagnosing or assessing disease severity.

[0010] The present invention provides applications of PRMT5, including applications of PRMT5 in the preparation of drugs or reagents for detecting or treating systemic sclerosis. Specifically, anti-PRMT5 antibodies can be used as markers for detecting systemic sclerosis. PRMT5 can be used to detect the level of anti-PRMT5 antibodies in a sample using an enzyme-linked immunosorbent assay (ELISA) to assist in determining the severity of systemic sclerosis medications or to indicate the progression and prognosis of skin and pulmonary fibrosis.

[0011] Preferably, anti-PRMT5 antibodies can be used to distinguish systemic sclerosis from healthy controls and other autoimmune disease patients. A preferred example of the present invention shows that the absorbance of the serum of SSc patients is significantly higher than that of HC (healthy controls), SLE patients and SjS patients, and anti-PRMT5 antibodies can be used as a specific biomarker for the diagnosis of SSc. Another preferred example of the present invention shows that the baseline level of anti-PRMT5 antibodies in the serum of SSc patients with progression of skin mRSS scores is significantly higher than that of patients without progression (Figure 2A); in patients with progression of mRSS scores, the level of anti-PRMT5 antibodies is significantly increased, while in patients with decreased mRSS scores, the level of anti-PRMT5 antibodies is significantly decreased. Anti-PRMT5 antibodies can also be used as an auxiliary to determine the outcome and prognosis of skin and pulmonary fibrosis.

[0012] Preferably, the level of anti-PRMT5 antibodies is measured by enzyme-linked immunosorbent assay, comprising the following steps:

[0013] S1, human PRMT5 protein was coated on a microplate and incubated overnight in a refrigerator;

[0014] S2, after incubation, wash with PBS containing TWEEN-20;

[0015] S3, add PBS containing bovine serum albumin to the microwells for blocking, and then add PBS for washing after incubation;

[0016] S4, dilute the serum 3-50 times using PBS containing BSA, add the diluted serum to the microplate, and incubate at 35-38°C for at least 0.5 h;

[0017] S5, aspirate the serum and add TBST to each well for washing;

[0018] S6, monoclonal anti-Fab-HRP antibody was added to each well and incubated;

[0019] S7, remove the antibody, add TBST to each well for washing, add the substrate ABTS to measure the enzyme activity, and read the signal using a microplate spectrophotometer;

[0020] S8, record the absorbance of serum at different dilution factors, plot the data and calculate the area under the curve (AUC).

[0021] In a preferred embodiment of the present invention, the enzyme-linked immunosorbent assay for determining the level of anti-PRMT5 antibodies comprises the following steps:

[0022] (1) 50 μl of 2 ng / μl recombinant human PRMT5 protein was coated on a 96-well microplate and incubated at 4°C overnight;

[0023] (2) After incubation, wash three times with 200 μl of PBS containing 0.1% (v / v) TWEEN-20;

[0024] (3) Add 100 μl of PBS containing 3% (w / v) bovine serum albumin to the microwells for blocking, incubate at 37°C for 1 h, and then wash three times with 100 μl of PBS;

[0025] (4) Serum was diluted 5-20 times using PBS containing 3% BSA, 100 μl of diluted serum was added to the microplate, and incubated at 37°C for 1.5 h;

[0026] (5) Aspirate the serum and add 100 μl of TBST to each well and wash three times;

[0027] (6) Add 50 μl of 5000-fold diluted monoclonal anti-Fab-HRP antibody to each well and incubate at 37°C for 45 min;

[0028] (7) The antibody was removed by aspiration, and 100 μl of TBST was added to each well for washing five times. The enzyme activity was determined by adding the substrate ABTS, and the signal was read at 405 nm using a microplate spectrophotometer;

[0029] (8) Record the absorbance of serum at different dilution factors, plot the data, and calculate the area under the curve (AUC). Use one-way ANOVA and Dunnett's multiple comparisons to analyze the differences in absorbance and AUC in serum of patients and healthy volunteers in each group;

[0030] (9) The anti-PRMT5 antibody level was higher than the upper limit of the 99% confidence interval of the HC group and was considered positive. The difference in the positive rate between SSc patients and HC was analyzed using Fisher's exact test;

[0031] (10) The receiver operating characteristic curve was drawn based on the absorbance of each group of samples, and the area under the curve was calculated.

[0032] The serum sample collection includes the following steps: collecting the sample, centrifuging at 200-500g for 3-10 minutes at 0-4°C, and aspirating the upper serum.

[0033] The anti-PRMT5 antibody content of the present invention can assist in determining the severity of systemic sclerosis, be used to indicate the progression and prognosis of skin and pulmonary fibrosis, and can also be corroborated with other detection methods. The detection methods that can be used include but are not limited to:

[0034] Patients were assessed based on the extent of skin involvement using the modified Rodnan skin score, which involves palpation of the skin at 17 anatomical sites to assess the extent and severity of skin thickening in SSc.

[0035] High-resolution computed tomography is performed to evaluate patients for interstitial lung disease:

[0036] Visual semiquantitative HRCT scoring was performed according to the type and degree of involvement of each lung region;

[0037] The Mann-Whitney U test was used to analyze the differences in baseline anti-PRMT5 antibody levels among different types of patients, and the paired t test was used to analyze the differences in changes in anti-PRMT5 antibody levels before and after follow-up among different types of patients;

[0038] Spearman correlation analysis was used to evaluate the correlation between anti-PRMT5 antibody levels and inflammation-related laboratory indicators in SSc patients.

[0039] Preferably, the sample is ex vivo serum from human peripheral blood.

[0040] In a preferred embodiment of the present invention, the level of anti-PMRT5 antibodies is used to diagnose diseases and indicate the progression and prognosis of skin and lung fibrosis.

[0041] The PRMT5 or anti-PRMT5 antibodies of the present invention can also be used to screen candidate drugs or agents for treating systemic sclerosis. After testing candidate materials for treating systemic sclerosis, the level of anti-PRMT5 antibodies in the sample is:

[0042] When the level of the anti-PRMT5 antibody in the sample is increased, the candidate material for treating systemic sclerosis is used as a candidate drug or agent for treating systemic sclerosis;

[0043] When the level of anti-PRMT5 antibodies in the sample is elevated, further testing of the candidate material for treating systemic sclerosis as a candidate drug or agent for treating systemic sclerosis is terminated.

[0044] Correspondingly, the present invention also provides a detection kit for systemic sclerosis, wherein the detection kit for systemic sclerosis contains PRMT5.

[0045] Preferably, the systemic sclerosis detection kit includes but is not limited to:

[0046] Anti-PRMT5 antibody diluent, anti-PRMT5 antibody buffer, PRMT5 quantitative standard, container for liquid containing PRMT5 or anti-PRMT5 antibody, PRMT5 antibody positive control, antibody negative control, detection consumables, deionized water.

[0047] The present invention provides a biomarker for systemic sclerosis, which can be used for the diagnosis and prognosis of the disease. The biomarker is an anti-PRMT5 antibody. The present invention is the first to discover that the serum level of anti-PRMT5 antibodies is elevated in systemic sclerosis patients, which can be used to distinguish systemic sclerosis from healthy controls and other autoimmune disease patients with good sensitivity and specificity. In addition, the level of anti-PRMT5 antibodies in systemic sclerosis patients is closely related to the progression of the patient's skin and pulmonary fibrosis, and has potential value in judging prognosis. The detection method of the present invention is an enzyme-linked immunosorbent assay, which is simple and easy to use, easy to transform and widely promoted in primary hospitals. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 shows the elevated levels of anti-PRMT5 antibodies in the serum of SSc patients.

[0049] 1A: When the serum is diluted 20 times, the enzyme-linked immunosorbent assay is used to detect the serum levels of SSc patients, systemic lupus erythematosus (SLE), and Sjögren's syndrome ( s syndrome (SjS) and healthy controls (HC); 1B: The absorbance at different dilution factors was plotted, and the area under the curve (AUC) was calculated. The AUC in SSc patients was significantly higher than that in HC, SLE, and SjS patients; 1C: The upper limit of the 99% confidence interval of the anti-PRMT5 antibody level in the serum of healthy controls was defined as the positive judgment value for the anti-PRMT5 antibody level, and the positive rate of anti-PRMT5 antibodies in the serum of SSc patients and HC was compared; 1D: The receiver operating characteristic (ROC) curve of the serum PRMT5 antibody levels of SSc patients was plotted to compare with those of the healthy control group, SLE group, and SjS group, and the area under the curve (AUC) was calculated.

[0050] Figure 2 is a correlation diagram between serum anti-PRMT5 antibody levels and SSc clinical characteristics in SSc patients, where

[0051] 2A: Comparison of anti-PRMT5 antibody levels between patients with mRSS score progression (≥25%) and those without progression within the past 12 ± 2 months; 2B: Comparison of changes in anti-PRMT5 antibody levels before and after follow-up in SSc patients with mRSS score progression or remission; 2C: Comparison of anti-PRMT5 antibody levels between patients with interstitial lung disease progression as shown by HRCT within the past 12 ± 2 months and those without progression; 2D: Comparison of changes in anti-PRMT5 antibody levels before and after follow-up in SSc patients with interstitial lung disease progression or remission as shown by HRCT. 2E: Correlation analysis of serum anti-PRMT5 antibody levels with erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin G (IgG), and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in SSc patients.

[0052] The following examples of the present application will clearly and completely describe the technical solution. Obviously, the described examples are only some preferred embodiments of the present application, not all embodiments. Based on the examples in the present application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application. DETAILED DESCRIPTION

[0053] The anti-PRMT5 biomarker antibodies of the present invention can be used to prepare a kit for SSc disease diagnosis and prognosis prediction. The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without inventive effort are within the scope of protection of the present invention.

[0054] Example 1 Determination of anti-PRMT5 antibody levels in serum of systemic sclerosis patients using enzyme-linked immunosorbent assay

[0055] The operation process is as follows:

[0056] 1. Serum sample collection

[0057] Peripheral blood was collected from 90 SSc patients, 30 SLE patients, 8 SjS patients and 84 sex- and age-matched healthy volunteers (HC). The blood was centrifuged at 300 g for 5 min at 4°C, and the upper serum was aspirated and immediately stored at -80°C.

[0058] 2. Enzyme-linked immunosorbent assay to determine anti-PRMT5 antibody levels

[0059] (1) 50 μl of 2 ng / μl recombinant human PRMT5 protein (Sino Biological, #11074-H18H) was coated on a 96-well microplate (Corning) and incubated at 4°C overnight.

[0060] (2) After incubation, wash three times with 200 μl of PBS containing 0.1% (v / v) TWEEN-20 (PBST);

[0061] (3) Add 100 μl of PBS containing 3% (w / v) bovine serum albumin (BSA) to the microwells for blocking. Incubate at 37°C for 1 h, then wash three times with 100 μl of PBS.

[0062] (4) Serum was diluted 5×, 10×, 20×, and 40× using PBS containing 3% BSA. 100 μl of the diluted serum was added to the microplate and incubated at 37°C for 1.5 h.

[0063] (5) Aspirate the serum and add 100 μl of TBST to each well and wash three times.

[0064] (6) Add 50 μl of 5000-fold diluted monoclonal anti-Fab-HRP antibody (Sigma-Aldrich) to each well and incubate at 37°C for 45 min.

[0065] (7) The antibody was removed by aspiration, and 100 μl of TBST was added to each well for washing five times. The enzyme activity was determined by adding the substrate ABTS (Invitrogen), and the signal was read at 405 nm using a microplate spectrophotometer (Biotek).

[0066] (8) The absorbance of serum at different dilution factors was recorded, the data were plotted, and the area under the curve (AUC) was calculated. One-way ANOVA and Dunnett's multiple comparisons were used to analyze the differences in absorbance and AUC in the serum of patients and healthy volunteers in each group.

[0067] (9) The anti-PRMT5 antibody level was higher than the upper limit of the 99% confidence interval (CI) of the HC group and was considered positive. The difference in the positive rate between SSc patients and HC was analyzed using Fisher's exact test.

[0068] (10) The receiver operating characteristic curve was drawn based on the absorbance of each group of samples, and the area under the curve was calculated.

[0069] 3. Results Analysis

[0070] (1) The results showed that when the serum was diluted 20 times, the absorbance of the serum of SSc patients was significantly higher than that of HC, SLE patients, and SjS patients (SSc vs HC p < 0.0001; SSc vs SLE p < 0.0001; SSc vs SjS p = 0.0030) (Figure 1A). In addition, the AUC of the serum of SSc patients was also significantly higher than that of HC, SLE, or SjS patients (SSc vs HC p < 0.0001; SSc vs SLE p < 0.0001; SSc vs SjS p = 0.0027) (Figure 1B). These results indicate that the level of anti-PRMT5 antibodies in the serum of SSc patients is significantly increased compared with healthy subjects or SLE and SjS patients.

[0071] (2) The 99th percentile of the HC group was used as the upper limit of normal to define the positive judgment value of the antibody. The results showed that 31.11% of SSc patients (28 / 90) had anti-PRMT5 antibodies, while no healthy controls (0 / 84) had anti-PRMT5 antibodies (Figure 1C). There was a significant difference between the two (p < 0.0001). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 70.24%, 97.78%, 96.72%, and 77.88%, respectively. In addition, anti-PRMT5 also showed the ability to distinguish SSc from SLE and SjS patients, with AUCs of 0.968 and 0.988, respectively (Figure 3D). Taken together, these results indicate that anti-PRMT5 antibodies can be used as a specific biomarker for the diagnosis of SSc.

[0072] Example 2 Analysis of the correlation between serum anti-PRMT5 antibody levels and SSc clinical characteristics in SSc patients

[0073] 1. Clinical follow-up of SSc patients:

[0074] (1) Eighteen SSc patients were followed up for 12±2 months during treatment with immunosuppressants and / or antifibrotic drugs. Clinical data, including clinical manifestations and laboratory parameters, were collected.

[0075] (2) According to the method in Example 1, the anti-PRMT5 antibody level was determined by enzyme-linked immunosorbent assay.

[0076] (3) The modified Rodnan skin score (mRSS) was performed on the patients according to the degree of skin involvement. The skin of 17 anatomical sites was palpated to assess the extent and severity of skin thickening in SSc.

[0077] (4) Perform high-resolution computed tomography (HRCT) to assess the patient's interstitial lung disease: if HRCT shows that the area affected by pulmonary fibrosis exceeds 10%, it is diagnosed as interstitial lung disease (ILD); perform visual semi-quantitative HRCT scoring based on the type and degree of involvement of each lung area.

[0078] (5) The Mann-Whitney U test was used to analyze the differences in baseline levels of anti-PRMT5 antibodies among different types of patients, and the paired t test was used to analyze the differences in changes in anti-PRMT5 antibody levels before and after follow-up among different types of patients.

[0079] (6) Spearman correlation analysis was used to evaluate the correlation between anti-PRMT5 antibody levels and inflammation-related laboratory indicators in SSc patients.

[0080] 2. Result analysis:

[0081] (1) The results showed that the baseline level of anti-PRMT5 antibodies in the serum of SSc patients with progressive skin mRSS scores was significantly higher than that of patients without progression (p = 0.0264) (Figure 2A); further follow-up of the changes in anti-PRMT5 antibody levels in the serum of SSc patients after 12 ± 2 months showed that the anti-PRMT5 antibody level was significantly increased in patients with progressive mRSS scores (p = 0.06), while the anti-PRMT5 antibody level was significantly decreased in patients with decreased mRSS scores (p = 0.002) (Figure 2B). The above results suggest that the level of anti-PRMT5 antibodies in the serum of SSc patients is correlated with skin fibrosis.

[0082] (2) The results showed that the baseline level of anti-PRMT5 antibodies in the serum of SSc patients with HRCT score progression was significantly higher than that of patients without progression (p = 0.0069) (Figure 2C); further follow-up of the changes in the anti-PRMT5 antibody levels in the serum of SSc patients after 12 ± 2 months showed that the anti-PRMT5 antibody levels were significantly increased in patients with HRCT score progression (p = 0.0088), while the anti-PRMT5 antibody levels were significantly decreased in patients with HRCT score improvement (p = 0.03) (Figure 2D). The above results suggest that the anti-PRMT5 antibody levels in the serum of SSc patients are correlated with pulmonary fibrosis.

[0083] (3) The results showed that the serum anti-PRMT5 antibody levels in SSc patients were positively correlated with ESR, CRP, IgG, and TIMP-1 levels (Figure 2E). The above results suggest that the serum anti-PRMT5 antibody levels in SSc patients may be correlated with the patients' inflammatory immune response levels.

[0084] In the description of the present invention, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0085] The embodiments described above are merely specific implementations of the present application, but the scope of protection of the present application is not limited thereto. Any changes or substitutions that can be imagined by any person skilled in the art within the technical scope disclosed in the present application without resorting to creative effort should be included within the scope of protection of the present application. Therefore, the scope of protection of the present application should be based on the scope of protection of the claims in the present application.

Claims

1. Application of PRMT5 as a biomarker in the preparation and detection of systemic sclerosis preparations.

2. The use according to claim 1, characterized in that Anti-PRMT5 antibodies are markers for detecting systemic sclerosis. PRMT5 is used to detect the level of anti-PRMT5 antibodies in samples using an enzyme-linked immunosorbent assay to assist in determining the severity of systemic sclerosis medication, distinguish systemic sclerosis from healthy controls and patients with other autoimmune diseases, or to indicate the progression and prognosis of skin and pulmonary fibrosis.

3. The use according to claim 2, characterized in that Anti-PRMT5 antibody levels were measured by enzyme-linked immunosorbent assay using the following steps: S1, human PRMT5 protein was coated on a microplate and incubated overnight in a refrigerator; S2, after incubation, wash with PBS containing TWEEN-20; S3, add PBS containing bovine serum albumin to the microwells for blocking, incubation and washing with PBS; S4, dilute the serum 3-50 times using PBS containing BSA, add the diluted serum to the microplate, and incubate at 35-38°C for at least 0.5 h; S5, aspirate the serum and add TBST to each well for washing; S6, monoclonal anti-Fab-HRP antibody was added to each well and incubated; S7, remove the antibody, add TBST to each well for washing, add the substrate ABTS to measure the enzyme activity, and read the signal using a microplate spectrophotometer; S8, record the absorbance of serum at different dilution factors, plot the data and calculate the area under the curve (AUC).

4. The use according to claim 3, characterized in that The enzyme-linked immunosorbent assay (ELISA) for measuring anti-PRMT5 antibody levels involves the following steps: (1) 50 μl of 2 ng / μl recombinant human PRMT5 protein was coated on a 96-well microplate and incubated at 4°C overnight; (2) After incubation, wash three times with 200 μl of PBS containing 0.1% (v / v) TWEEN-20; (3) Add 100 μl of PBS containing 3% (w / v) bovine serum albumin to the microwells for blocking, incubate at 37°C for 1 h, and then wash three times with 100 μl of PBS; (4) Use PBS containing 3% BSA to dilute the serum 5-20 times, and add 100 μl of the diluted serum to Place in a microplate and incubate at 37°C for 1.5 h; (5) Aspirate the serum and add 100 μl of TBST to each well and wash three times; (6) Add 50 μl of 5000-fold diluted monoclonal anti-Fab-HRP antibody to each well and incubate at 37°C for 45 min; (7) The antibody was removed by aspiration, and 100 μl of TBST was added to each well for washing five times. The enzyme activity was determined by adding the substrate ABTS, and the signal was read at 405 nm using a microplate spectrophotometer; (8) Record the absorbance of serum at different dilution factors, plot the data, and calculate the area under the curve (AUC). Use one-way ANOVA and Dunnett's multiple comparisons to analyze the differences in absorbance and AUC in serum of patients and healthy volunteers in each group; (9) The anti-PRMT5 antibody level was higher than the upper limit of the 99% confidence interval of the HC group and was considered positive. The difference in the positive rate between SSc patients and HC was analyzed using Fisher's exact test; (10) Draw the receiver operating characteristic curve based on the absorbance of each group of samples and calculate the area under the curve; and / or The serum sample collection includes the following steps: collecting the sample, centrifuging at 200-500g for 3-10 minutes at 0-4°C, and aspirating the upper serum.

5. The use according to claim 1, characterized in that The application adopts one or more of the following method steps: Patients were assessed based on the extent of skin involvement using the modified Rodnan skin score, which involves palpation of the skin at 17 anatomical sites to assess the extent and severity of skin thickening in SSc. High-resolution computed tomography is performed to evaluate patients for interstitial lung disease: Visual semiquantitative HRCT scoring was performed according to the type and degree of involvement of each lung region; The Mann-Whitney U test was used to analyze the differences in baseline anti-PRMT5 antibody levels among different types of patients, and the paired t test was used to analyze the differences in changes in anti-PRMT5 antibody levels before and after follow-up among different types of patients; Spearman correlation analysis was used to evaluate the correlation between anti-PRMT5 antibody levels and inflammation-related laboratory indicators in SSc patients.

6. The use according to any one of claims 1 to 5, characterized in that The sample is ex vivo serum from human peripheral blood.

7. The use according to claim 1, characterized in that Application of PRMT5 in screening candidate drugs or reagents for the treatment of systemic sclerosis: Detecting the level of anti-PRMT5 antibodies in samples after the use of candidate materials for the treatment of systemic sclerosis; When the level of the anti-PRMT5 antibody in the sample is reduced, the candidate material for treating systemic sclerosis is used as a candidate drug or agent for treating systemic sclerosis; When the level of anti-PRMT5 antibodies in the sample is elevated, further testing of the candidate material for treating systemic sclerosis as a candidate drug or agent for treating systemic sclerosis is terminated.

8. A detection kit for systemic sclerosis, characterized in that: The systemic sclerosis detection kit contains PRMT5 or anti-PRMT5 antibody.

9. The detection kit for systemic sclerosis according to claim 8, characterized in that The systemic sclerosis detection kit contains one or more of the following: Anti-PRMT5 antibody diluent, anti-PRMT5 antibody buffer, PRMT5 quantitative standard, container for liquid containing PRMT5 or anti-PRMT5 antibody, PRMT5 or anti-PRMT5 antibody positive control, PRMT5 or anti-PRMT5 antibody negative control, detection consumables, deionized water.

Images