Application of TRIM69 in preparation of kit for auxiliary diagnosis of systemic lupus erythematosus

Abstract

The invention relates to the technical field of biological medicine, and provides application of TRIM69 in preparation of a kit for auxiliary diagnosis of systemic lupus erythematosus. The kit assists in diagnosing systemic lupus erythematosus by detecting the TRIM69 level. Through the mRNA expression quantity of the TRIM69 and the expression quantity and / or concentration of the protein, the auxiliary diagnosis of the systemic lupus erythematosus is realized, and the TRIM69 as a biomarker has important values in the aspects of auxiliary diagnosis, disease activity monitoring, prognosis evaluation, diagnosis efficiency improvement and the like. Not only can a doctor be helped to recognize a high-risk patient earlier, but also a basis can be provided for individualized treatment, so that the long-term prognosis of the systemic lupus erythematosus patient is improved.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of TRIM69 in preparing a kit for auxiliary diagnosis of systemic lupus erythematosus. Background Art

[0002] Systemic lupus erythematosus (SLE) is an autoimmune disease affecting multiple systems and organs. Its pathogenesis is complex, its clinical manifestations are highly heterogeneous, and its early symptoms can be easily confused with other diseases, such as rheumatoid arthritis and Sjögren's syndrome. Therefore, accurate diagnosis of SLE is crucial for controlling disease progression, reducing the risk of organ damage, and improving patient prognosis.

[0003] Currently, the diagnosis of systemic lupus erythematosus (SLE) relies primarily on a combination of clinical symptoms, laboratory tests, and imaging assessments. However, this process lacks specificity, is time-consuming, and can lead to irreversible organ damage. Commonly used immunological markers for SLE have numerous limitations, such as a high false-positive rate for ANA and low sensitivity for anti-dsDNA and anti-Sm antibodies, which can lead to missed diagnoses in some cases. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention provides the use of TRIM69 in the preparation of a kit for auxiliary diagnosis of systemic lupus erythematosus, which aims to solve the problems mentioned in the background technology.

[0005] In a first aspect, the present invention provides a use of TRIM69 in preparing a kit for auxiliary diagnosis of systemic lupus erythematosus, wherein the kit assists in the diagnosis of systemic lupus erythematosus by detecting the level of TRIM69.

[0006] Furthermore, the kit assists in the diagnosis of systemic lupus erythematosus by detecting the mRNA expression level, protein expression level and / or concentration of TRIM69.

[0007] Furthermore, the kit includes an antibody for detecting TRIM69 protein or a PCR primer for the cDNA chain of the TRIM69 gene.

[0008] Furthermore, the biological sample detected by the kit includes peripheral blood or serum.

[0009] Furthermore, TRIM69 mRNA expression was elevated in the peripheral blood of patients with systemic lupus erythematosus; The expression level and / or concentration of TRIM69 protein in the serum of patients with systemic lupus erythematosus is increased.

[0010] Furthermore, when the concentration of TRIM69 protein in the serum of the subject detected by the ELISA method is greater than 187.2 pg / ml, the kit assists in diagnosing that the subject is a patient with systemic lupus erythematosus, wherein the subject does not include patients with rheumatoid arthritis.

[0011] In a second aspect, the present invention provides a prediction model for diagnosing systemic lupus erythematosus based on serum TRIM69 levels, hemoglobin, and neutrophil-to-lymphocyte ratio, wherein the prediction model formula is expressed as: ; Where: Q is the predicted value, HGB is hemoglobin, NLR is the neutrophil-to-lymphocyte ratio, and TRIM69 is the protein expression level.

[0012] Furthermore, when the predicted value is greater than 0.324, the subject is diagnosed as a patient with systemic lupus erythematosus, wherein the subject does not include patients with rheumatoid arthritis.

[0013] Furthermore, when the predicted value is greater than 1.558, the subject is auxiliary diagnosed as a patient with systemic lupus erythematosus.

[0014] The present invention has the following beneficial effects: (1) TRIM69 is used in the preparation of auxiliary diagnosis kits for systemic lupus erythematosus. By measuring the mRNA expression level, protein expression level and / or concentration of TRIM69, the diagnosis of systemic lupus erythematosus can be assisted. As a biomarker, TRIM69 has important value in auxiliary diagnosis, disease activity monitoring, prognosis assessment and improving diagnostic efficiency. It can not only help doctors identify high-risk patients earlier, but also provide a basis for individualized treatment, thereby improving the long-term prognosis of patients with systemic lupus erythematosus. With the development of more research, TRIM69 is expected to become a key tool in the management of systemic lupus erythematosus.

[0015] (2) The prediction model constructed for diagnosing systemic lupus erythematosus based on serum TRIM69 levels, hemoglobin, and neutrophil-to-lymphocyte ratio can more accurately identify true systemic lupus erythematosus patients (high sensitivity) and effectively exclude non-patients (high specificity), reducing the risk of misdiagnosis and missed diagnosis, and providing clinicians with a more reliable diagnostic basis. In addition, compared with a single indicator, the multi-element prediction model has stronger comprehensive diagnostic capabilities and can integrate information provided by different indicators, thereby overcoming the limitations of a single indicator (such as large volatility, insufficient specificity, etc.) and improving the stability and accuracy of the overall prediction. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] A more complete understanding of exemplary embodiments of the present invention may be obtained by referring to the following drawings: Figure 1 The RT-qPCR test results and ELISA test results of Example 2 of the present invention are as follows: Figure 1 Figure A is a statistical graph of the RT-qPCR detection results of TRIM69 expression in the peripheral blood of patients with systemic lupus erythematosus, rheumatoid arthritis, and healthy controls. GADPH was used as an internal reference. *** indicates p < 0.001, **** indicates p < 0.0001. Figure 1 Figure B is a statistical graph of the RT-qPCR detection results of TRIM69 expression in the peripheral blood of the systemic lupus erythematosus patient group and the rheumatoid arthritis patient + healthy control group. GADPH was used as an internal reference. **** indicates p < 0.0001; Figure 1 Figure C is a statistical graph of serum TRIM69 protein concentrations in the systemic lupus erythematosus patient group, rheumatoid arthritis patient group, and healthy control group. ** indicates p < 0.01, and **** indicates p < 0.0001. Figure 1 D in the figure is a statistical graph of TRIM69 protein concentration in the serum of the systemic lupus erythematosus patient group and the rheumatoid arthritis patient + healthy control group, *** indicates p < 0.001.

[0017] Figure 2 3 is a graph showing the correlation analysis results between the serum TRIM69 level in systemic lupus erythematosus patients and the clinical characteristics of systemic lupus erythematosus patients according to Example 3 of the present invention, wherein: Figure 2 A in the figure is the correlation analysis result between serum TRIM69 protein concentration and white blood cell count, p < 0.0001; Figure 2 Figure B is the correlation analysis result between serum TRIM69 protein concentration and lymphocyte absolute value, p < 0.0001; Figure 2 Figure C is the correlation analysis result between serum TRIM69 protein concentration and monocyte absolute value, p < 0.0001; Figure 2 Figure D is the correlation analysis result between serum TRIM69 protein concentration and absolute number of neutrophils, p < 0.0001; Figure 2 Figure E is the correlation analysis result between serum TRIM69 protein concentration and platelet-to-lymphocyte ratio, p < 0.0001; Figure 2 Figure F is the correlation analysis result between serum TRIM69 protein concentration and complement C3, p < 0.0001; Figure 2 G in the figure is the statistical graph of TRIM69 protein concentration in the serum of patients with anti-SSA-positive and anti-SSA-negative systemic lupus erythematosus, p=0.0181; Figure 2 H in the figure is the statistical graph of TRIM69 protein concentration in the serum of anti-Ro52-positive and anti-Ro52-negative systemic lupus erythematosus patients, p=0.0165; Figure 2 Figure I is a statistical graph of TRIM69 protein concentration in the serum of anti-Sm-positive and anti-Sm-negative systemic lupus erythematosus patients, p=0.0149.

[0018] Figure 3 : is the ROC curve analysis result diagram of Example 4 of the present invention, wherein: Figure 3 Figure A is the ROC curve analysis result of the serum TRIM69 level in patients with systemic lupus erythematosus and healthy controls, P < 0.0001; Figure 3 Figure B is the ROC curve analysis result of the TRIM69 level in the serum of patients with systemic lupus erythematosus and rheumatoid arthritis patients + healthy controls, P < 0.0001.

[0019] Figure 4 is the analysis result of the prediction model constructed by the serum TRIM69 level, hemoglobin, and neutrophil-to-lymphocyte ratio in Example 5 of the present invention, wherein: Figure 4 A in the figure is a statistical diagram of the predicted values ​​of the systemic lupus erythematosus patient group and the healthy control group, p < 0.0001; Figure 4 Figure B is the ROC analysis of the prediction model, which distinguished patients with systemic lupus erythematosus from healthy controls, P < 0.0001; Figure 4 C in the figure is a statistical graph of the predicted values ​​of the systemic lupus erythematosus patient group and the rheumatoid arthritis patient + healthy control group, p < 0.0001; Figure 4 D in the figure is the ROC analysis of the prediction model, which distinguishes patients with systemic lupus erythematosus from patients with rheumatoid arthritis and healthy controls, P < 0.0001.

[0020] Figure 5 is the analysis result of the prediction model of serum TRIM69 level, red blood cell count, and lymphocyte-to-monocyte ratio in Example 5 of the present invention, wherein: Figure 5 A in the figure is a statistical diagram of the predicted values ​​of the systemic lupus erythematosus patient group and the healthy control group, p=0.0359; Figure 5 Figure B is a statistical graph of the predicted values ​​of the systemic lupus erythematosus patient group and the rheumatoid arthritis patient + healthy control group, p=0.9196; Figure 5 C in the figure is the ROC analysis of the prediction model, which distinguishes patients with systemic lupus erythematosus from healthy controls with P=0.036.

[0021] Figure 6 : This is a graph showing the analysis results of the relationship between the prediction model constructed using serum TRIM69 levels, hemoglobin, and neutrophil-to-lymphocyte ratio in Example 6 of the present invention and the clinical characteristics of patients with systemic lupus erythematosus, wherein: Figure 6 A in the figure is the correlation analysis result between the prediction model and the percentage of neutrophils, p=0.0001; Figure 6 Figure B is the correlation analysis result between the prediction model and the platelet-to-lymphocyte ratio, p < 0.0001; Figure 6 C in the figure is the correlation analysis result between the prediction model and the neutrophil-to-lymphocyte ratio, p < 0.0001; Figure 6 D in the figure is the correlation analysis result between the prediction model and red blood cell count, p=0.0003; Figure 6 E in the figure is the correlation analysis result between the prediction model and hemoglobin percentage, p < 0.0001; Figure 6 F in the figure is the correlation analysis result between the prediction model and hematocrit, p < 0.0001; Figure 6 G in the figure is the correlation analysis result between the prediction model and the absolute value of lymphocytes, p < 0.0001; Figure 6 H in the figure is the correlation analysis result between the prediction model and the percentage of lymphocytes, p < 0.0001; Figure 6 I in the figure is the correlation analysis result between the prediction model and the lymphocyte-to-monocyte ratio, p=0.0044. DETAILED DESCRIPTION

[0022] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; the terms used herein are only for the purpose of describing specific embodiments rather than limiting the present invention.

[0024] An embodiment of the present invention provides the use of TRIM69 in preparing a kit for auxiliary diagnosis of systemic lupus erythematosus, wherein the kit assists in the diagnosis of systemic lupus erythematosus by detecting the level of TRIM69.

[0025] In some embodiments, the kit assists in the diagnosis of systemic lupus erythematosus by detecting the mRNA expression level, protein expression level and / or concentration of TRIM69.

[0026] In some embodiments, the kit includes an antibody for detecting TRIM69 protein or a PCR primer for detecting the cDNA strand of the TRIM69 gene.

[0027] In some embodiments, the biological sample detected by the kit includes peripheral blood or serum.

[0028] In some embodiments, the mRNA expression level of TRIM69 in the peripheral blood of patients with systemic lupus erythematosus is elevated; The expression level and / or concentration of TRIM69 protein in the serum of patients with systemic lupus erythematosus is increased.

[0029] In some embodiments, when the concentration of TRIM69 protein in the serum of the subject detected by the kit through ELISA is greater than 187.2 pg / ml, the subject is diagnosable as a patient with systemic lupus erythematosus, wherein the subject does not include patients with rheumatoid arthritis.

[0030] In some embodiments, the present invention provides a prediction model for diagnosing systemic lupus erythematosus based on serum TRIM69 levels, hemoglobin, and neutrophil-to-lymphocyte ratio. The prediction model formula is expressed as: ; Where: Q is the predicted value, HGB is hemoglobin, NLR is the neutrophil-to-lymphocyte ratio, and TRIM69 is the protein expression level.

[0031] In some embodiments, the higher the predicted value, the higher the risk of systemic lupus erythematosus.

[0032] In some embodiments, when the prediction value is greater than 0.324, the subject is diagnosed as a patient with systemic lupus erythematosus, wherein the subject does not include a patient with rheumatoid arthritis.

[0033] In some embodiments, when the prediction value is greater than 1.558, the subject is diagnosed as a patient with systemic lupus erythematosus.

[0034] Sample collection and analysis: (1) Subjects: From June 2024 to March 2025, patients with systemic lupus erythematosus (SLE) who met the revised diagnostic criteria of the American College of Rheumatology (ACR) for SLE were recruited at the First Affiliated Hospital of Nanchang University. Healthy controls (without autoimmune diseases or other inflammatory diseases) matched for age and sex were also selected. The disease activity of SLE patients was assessed using the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), and patients were divided into stable phase (SLEDAI 0-9 points) and active phase (SLEDAI ≥ 10 points) based on the score. Among the SLE patients, 9 newly diagnosed patients had not received immunosuppressive drugs or corticosteroids before enrollment, and the remaining patients had been previously diagnosed, treated, and relapsed. At the same time, patients with rheumatoid arthritis (RA) admitted to the hospital were included as disease controls. All patients with RA met the revised diagnostic criteria of the American College of Rheumatology for RA. The protocol was approved by the Ethics Committee of the First Affiliated Hospital of Nanchang University, approval number: (2023)CDYFYYLK(01-002), and all participants signed written informed consent.

[0035] (2) RNA extraction: EDTA-anticoagulated serum was obtained from the subjects for peripheral blood collection. RNA was extracted from whole blood using TRIzol® (Thermo Fisher Scientific) according to the manufacturer's protocol. RNA concentration was quantified and identified using a NanoDrop ND-1000 spectrophotometer (Agilent Technologies) and stored at −80°C.

[0036] (3) RT-qPCR (reverse transcription-quantitative polymerase chain reaction) analysis: Total RNA was reverse transcribed into cDNA using a reverse transcription kit (Tara Biotech Co., Ltd.). qPCR amplification of the cDNA samples was performed on an ABI 7500 real-time PCR instrument (Applied Biosystems; Thermo Fisher Scientific). The primers for TRIM69 were: forward primer: 5'-CTTGCCATCCAACAGGGTCAA-3' (SEQ ID NO. 1), reverse primer: 5'-TTCCTTGTGAGCAGCAATAGC-3' (SEQ ID NO. 2), designed using Primers 5 software, verified by primer BLAST, and synthesized by Shanghai Shenggong. In each sample, GADPH was used as an internal control, forward primer: 5'-TGCACCACCAACTGCTTAGC-3' (SEQ ID NO. 3), reverse primer: 5'-GGCATGGACTGTGGTCATGAG-3' (SEQ ID NO. 4). All RT-qPCR data were analyzed using the 2-∆∆Ct method.

[0037] (4) ELISA (enzyme-linked immunosorbent assay): Whole blood samples were collected and centrifuged (3500 rpm for 10 minutes) to obtain serum, which was then stored at -80°C. The concentration of TRIM69 protein in the serum was determined using an ELISA kit (Abesa, abx383943).

[0038] (5) Routine blood tests, serum inflammatory markers, and autoantibody testing: Routine blood tests and urine tests were performed using the Thermo Fisher X2100 (Thermo Fisher Scientific, Hanshin, Japan) and Thermo Fisher UF1000i (Thermo Fisher Scientific, Japan). Serum C3 (complement 3), C4 (complement 4), CRP (C-reactive protein), and IgG (immunoglobulin G) levels were determined by turbidimetric methods using the Immuno800 system (Beckman Coulter). ESR (erythrocyte sedimentation rate) was determined according to the manufacturer's instructions. IgG anti-dsDNA in serum was detected using an ELISA kit (Shanghai Kexin Co., Ltd.), and ANA (antinuclear antibody) was determined using an indirect immunofluorescence assay (Oremin). Linear immunoassay kits (Oermin, Lübeck, Germany) were used to measure anti-ENA (anti-extractable nuclear antigen antibodies), including anti-SSA (anti-Sjögren's syndrome A antigen antibody), anti-SSB (anti-Sjögren's syndrome B antigen antibody), anti-Ro52 (anti-trimeric protein 21 antibody), anti-Sm (anti-Smith antibody), anti-PO (anti-phosphatase kinase antibody), anti-U1-snRNP (anti-U1 small nuclear ribonucleoprotein antibody), anti-ANuA (anti-antinuclear antibody), and anti-Histone (anti-histone antibody).

[0039] (6) Statistical analysis: All data are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using Graphpad Prism 10.1.2 software. For comparisons between two groups, Student's t-test or Mann-Whitney U test was used, depending on whether the data conformed to a normal distribution. For comparisons between three groups, the Kruskal-Wallis test was used, followed by Dunn's post hoc test. Correlation analysis was performed using the Pearson or Spearman method. The receiver operating characteristic (ROC) curve was used to evaluate the predictive performance of TRIM69, routine blood parameters, and the prediction model.

[0040] Example 1: Detailed demographic characteristics of 199 participants (72 patients with systemic lupus erythematosus, 62 healthy controls, and 65 patients with rheumatoid arthritis) were collected.

[0041] The statistical results are shown in Table 1. Compared with healthy controls, patients with systemic lupus erythematosus exhibited significantly increased platelet distribution width (PDW), monocyte count and percentage (M, M%), and neutrophil count and percentage (N, N%), while red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean platelet volume (MPV), and lymphocyte count and percentage (L, L%) were significantly decreased. Compared with patients with rheumatoid arthritis, patients with systemic lupus erythematosus had significantly increased monocyte count and percentage (M, M%), while hemoglobin (HGB), hematocrit (HCT), platelet count (PLT), and platelet volume (PCT) were significantly decreased.

[0042] Table 1 Clinical characteristics of patients with systemic lupus erythematosus, healthy subjects, and rheumatoid arthritis

[0043] In the table, * indicates systemic lupus erythematosus versus healthy control group, p < 0.05; # indicates systemic lupus erythematosus versus rheumatoid arthritis group, p < 0.05; age, systemic lupus erythematosus disease activity score, erythrocyte sedimentation rate, hemoglobin, and platelet data have been rounded to two decimal places.

[0044] Example 2: (1) The mRNA levels of TRIM69 in the peripheral blood of patients with systemic lupus erythematosus, rheumatoid arthritis and healthy controls were compared by RT-qPCR.

[0045] RT-qPCR test results are as follows Figure 1 As shown in A and B, the results showed that the mRNA expression of TRIM69 in the peripheral blood of the systemic lupus erythematosus patient group was significantly upregulated, not only significantly higher than that of the healthy control group, but also significantly higher than that of the rheumatoid arthritis patient group ( Figure 1 In addition, the mRNA level of TRIM69 in the peripheral blood of the SLE group was significantly higher than that of the SLE group plus the healthy control group (Figure 1, A).

[0046] (2) Since the significant differences in mRNA levels were mainly observed in patients with systemic lupus erythematosus, the TRIM69 level (protein) in the serum of patients with systemic lupus erythematosus was detected by ELISA (enzyme-linked immunosorbent assay).

[0047] ELISA test results Figure 1As shown in Figures C and D, the results showed that compared with the healthy control group and the rheumatoid arthritis patient group, the serum TRIM69 level in the SLE patient group was also significantly elevated, with specific values ​​of 221.14±126.81 pg / ml in the SLE patient group, 142.32±78.42 pg / ml in the healthy control group, and 167.95±109.30 pg / ml in the rheumatoid arthritis patient group (Figure 1, C). In addition, the serum TRIM69 level in the SLE patient group was significantly higher than that in the rheumatoid arthritis patient and healthy control group (Figure 1, D).

[0048] Example 3: To analyze the correlation between serum TRIM69 levels (protein) in patients with systemic lupus erythematosus and the clinical characteristics of patients with systemic lupus erythematosus.

[0049] The results of the correlation analysis between the serum TRIM69 level in patients with systemic lupus erythematosus and the clinical characteristics of patients with systemic lupus erythematosus are as follows Figure 2 The results showed that the level of TRIM69 in serum was negatively correlated with the counts of various immune cells, including total white blood cell count (r=-0.5729, Figure 2A), lymphocytes (r=-0.4866, Figure 2B), monocytes (r=-0.4995, Figure 2C), and neutrophils (r=-0.5328, Figure 2 Notably, serum TRIM69 levels were positively correlated with the inflammation-related platelet-lymphocyte ratio (PLR) (r=0.5931, Figure 2E). Regarding complement, serum TRIM69 levels were negatively correlated with C3 (r=-0.7829, Figure 2F).

[0050] Furthermore, serum TRIM69 levels were elevated in patients with specific autoantibody-positive SLE. Specifically, TRIM69 levels were significantly higher in patients with anti-SSA-positive SLE (Figure 2, G), anti-Ro52-positive SLE (Figure 2, H), and anti-Sm-positive SLE (Figure 2, I) than in patients with negative results for the corresponding antibodies. This suggests that TRIM69 has the potential to serve as an indicator for assessing the activity and severity of SLE.

[0051] Example 4: The serum TRIM69 level (protein) in patients with systemic lupus erythematosus, rheumatoid arthritis and healthy controls was detected by ELISA. The receiver operating characteristic (ROC) curve was drawn, and the area under the curve (AUC), 95% confidence interval (CI), sensitivity and specificity were calculated.

[0052] The ROC curve analysis results are as follows Figure 3 The results showed that serum TRIM69 levels have potential value in distinguishing patients with systemic lupus erythematosus from healthy controls, with an AUC of 0.787 and a 95% confidence interval of 0.628-0.842. Graphpad Prism 10.1.2 software analysis showed that the optimal cutoff value was 187.2 pg / ml, corresponding to a sensitivity of 75.00% and a specificity of 70.83% (Figure 3A). This means that when the serum TRIM69 protein concentration is >187.2 pg / ml, it can assist in the diagnosis of systemic lupus erythematosus. Furthermore, the serum TRIM69 level in patients with systemic lupus erythematosus was significantly higher than that in healthy controls and patients with rheumatoid arthritis. Based on this, a risk score analysis using TRIM69 was performed comparing patients with systemic lupus erythematosus (SLE) with patients with rheumatoid arthritis and healthy controls. The AUC was 0.739, with a 95% confidence interval of 0.602-0.763, a sensitivity of 68.80%, and a specificity of 70.83% (Figure 3, B). This demonstrates that TRIM69, as a diagnostic biomarker for SLE, possesses the sensitivity and specificity required for clinical auxiliary diagnosis.

[0053] Example 5: The serum TRIM69 level (protein) was combined with the conventional clinical indicators hemoglobin, neutrophil-to-lymphocyte ratio, and lymphocyte-to-monocyte ratio to construct a prediction model, which was analyzed by univariate and multivariate analysis, as shown in Table 2. The serum TRIM69 level was combined with the red blood cell count and lymphocyte-to-monocyte ratio to construct a prediction model, which was analyzed by univariate and multivariate analysis, as shown in Table 3.

[0054] Table 2 Univariate and multivariate analysis of risk factors associated with systemic lupus erythematosus1

[0055] Table 3 Univariate and multivariate analysis of risk factors associated with systemic lupus erythematosus

[0056] The serum TRIM69 level, hemoglobin, and neutrophil-to-lymphocyte ratio were determined to be the optimal model variables, and the prediction model formula established was expressed as follows: , the predicted value Q was calculated, where Q is the predicted value, HGB is hemoglobin (g / L), NLR is the neutrophil-to-lymphocyte ratio, and TRIM69 is the protein expression level (pg / mL).

[0057] The analysis results of the prediction model constructed by serum TRIM69 level, hemoglobin and neutrophil to lymphocyte ratio are as follows Figure 4 The results showed that the higher the predicted value, the higher the risk of systemic lupus erythematosus ( Figure 4 The prediction model performed well in distinguishing patients with systemic lupus erythematosus from healthy controls, with an AUC value of 0.941 and a 95% confidence interval of 0.846-0.975, which was significantly better than a single indicator ( Figure 4 B). It is worth noting that, after analysis by Graphpad prism 10.1.2 software, the optimal cutoff value of the prediction model was 0.324, which had high specificity (81.94%) and high sensitivity (93.55%). That is, when the prediction value was > 0.324, the subject could be diagnosed as a patient with systemic lupus erythematosus. Similarly, the prediction model could distinguish patients with systemic lupus erythematosus from patients with rheumatoid arthritis and healthy controls ( Figure 4 C in the figure), the AUC value was 0.805, and the 95% confidence interval was 0.691-0.836. After analysis by Graphpad Prism 10.1.2 software, the optimal cutoff value was 1.558, corresponding to a sensitivity of 77.17% and a specificity of 72.22% ( Figure 4 D in the figure), that is, when the predicted value is >1.558, it can assist in diagnosing the subject as a patient with systemic lupus erythematosus.

[0058] The serum TRIM69 level, red blood cell count, and lymphocyte-to-monocyte ratio were used to construct a disease prediction model, such as Figure 5 The results showed that there was a statistically significant difference in the predicted values ​​between patients with systemic lupus erythematosus and healthy controls ( Figure 5 A in ), but could not distinguish patients with systemic lupus erythematosus from patients with rheumatoid arthritis plus healthy controls ( Figure 5 B in ); and performed moderately well in distinguishing patients with systemic lupus erythematosus from healthy controls, with an AUC of 0.605 and a 95% confidence interval of 0.510-0.701 ( Figure 5 C), analyzed by Graphpad Prism 10.1.2 software, with a cutoff value of 5.212, the sensitivity was high (85.48%) but the specificity was low (38.89%).

[0059] Example 6: The relationship between a prediction model constructed from serum TRIM69 levels, hemoglobin, and the neutrophil-to-lymphocyte ratio and the clinical characteristics of patients with systemic lupus erythematosus was analyzed. The analysis included multiple indicators, including the Systemic Lupus Erythematosus Disease Activity Index, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and complete blood count.

[0060] The results of the analysis of the relationship between the prediction model constructed by serum TRIM69 level, hemoglobin and neutrophil-to-lymphocyte ratio and the clinical characteristics of patients with systemic lupus erythematosus are as follows Figure 6 The results showed that the predicted value calculated by the prediction model was significantly correlated with the disease activity of systemic lupus erythematosus. The specific correlations were as follows: the predicted value was positively correlated with the percentage of neutrophils (r=0.4325, Figure 6A), the predicted value was positively correlated with the platelet-to-lymphocyte ratio (r=0.5555, Figure 6B), and the predicted value was strongly positively correlated with the neutrophil-to-lymphocyte ratio (r=0.7229, Figure 6C). In addition, the predicted value was negatively correlated with red blood cells (r=-0.418, Figure 6D), the predicted value was negatively correlated with hemoglobin (r=-0.5374, Figure 6E), the predicted value was negatively correlated with hematocrit (r=-0.5243, Figure 6F), the predicted value was negatively correlated with lymphocytes (r=-0.592, Figure 6G), and the predicted value was negatively correlated with the percentage of lymphocytes (r=-0.6205, Figure 6D). Figure 6 The predicted value was negatively correlated with the lymphocyte-to-monocyte ratio (r=-0.3295, Figure 6, I). This suggests that the predictive model constructed using serum TRIM69 levels, hemoglobin, and the neutrophil-to-lymphocyte ratio has utility in assessing SLE disease activity.

[0061] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. The use of TRIM69 in the preparation of a kit for auxiliary diagnosis of systemic lupus erythematosus, characterized in that: The kit assists in the diagnosis of systemic lupus erythematosus by detecting the level of TRIM69.

2. The use according to claim 1, characterized in that: The kit assists in the diagnosis of systemic lupus erythematosus by detecting the mRNA expression level, protein expression level and / or concentration of TRIM69.

3. The use according to claim 2, characterized in that: The kit includes an antibody for detecting TRIM69 protein or a PCR primer for detecting the cDNA chain of the TRIM69 gene.

4. The use according to claim 3, characterized in that: The biological samples detected by the kit include peripheral blood or serum.

5. The use according to claim 4, characterized in that: TRIM69 mRNA expression is elevated in the peripheral blood of patients with systemic lupus erythematosus; The expression level and / or concentration of TRIM69 protein in the serum of patients with systemic lupus erythematosus is increased.

6. The use according to claim 5, characterized in that: When the concentration of TRIM69 protein in the serum of the subject detected by the kit through ELISA is greater than 187.2 pg / ml, the subject is assisted in diagnosing that the subject is a patient with systemic lupus erythematosus, wherein the subject does not include patients with rheumatoid arthritis.

7. A predictive model for diagnosing systemic lupus erythematosus based on serum TRIM69 levels, hemoglobin, and neutrophil-to-lymphocyte ratio, characterized by: The prediction model formula is expressed as: Q = 4.375-0.0818×HGB+1.441×NLR+0.015×TRIM69; Where: Q is the predicted value, HGB is hemoglobin, NLR is the neutrophil-to-lymphocyte ratio, and TRIM69 is the protein expression level.

8. The prediction model according to claim 7, wherein: The higher the predicted value, the higher the risk of systemic lupus erythematosus.

9. The prediction model according to claim 7, wherein: When the predicted value is greater than 0.324, the subject is diagnosed as a patient with systemic lupus erythematosus, wherein the subject does not include patients with rheumatoid arthritis.

10. The prediction model according to claim 7, wherein: When the predicted value is greater than 1.558, the subject is auxiliary diagnosed as a patient with systemic lupus erythematosus.

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