Systemic lupus erythematosus molecular diagnostic markers based on neutrophil characteristics

Abstract

The application discloses a molecular diagnostic marker for systemic lupus erythematosus based on neutrophil characteristics. The application provides application of a substance for detecting expression amounts of IFIH1 genes and / or CFLAR genes in preparation of products for assisting in diagnosis or screening or auxiliary screening of systemic lupus erythematosus. The application also provides application of a substance for detecting relative expression amounts of IFIH1 genes and CFLAR genes and a carrier loaded with a combined index C value model formula in preparation of products for assisting in diagnosis or screening or auxiliary screening of systemic lupus erythematosus. The application focuses on unique changes of neutrophils in SLE, selects neutrophil-specific gene expression markers IFIH1 and / or CFLAR, assists in early differential diagnosis of SLE, and constructs a combined diagnosis model of IFIH1 and CFLAR, thereby realizing diagnosis or screening of SLE, and having high sensitivity and specificity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and relates to a molecular diagnostic marker for systemic lupus erythematosus based on neutrophil signatures. BACKGROUND

[0002] Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by immune tolerance disorders, autoantibody production, immune complex formation and deposition, leading to tissue and organ damage. The incidence of SLE is gradually increasing. There are more than 1 million SLE patients in China, mainly in women of childbearing age. SLE has a low clinical remission rate, a high disability rate and a high mortality rate, with a 15-year cumulative mortality rate of up to 20%, thus causing a serious economic burden to families and society. SLE has diverse clinical phenotypes, multiple and severe organ involvement, often involving the kidney, cardiovascular system, skin, central nervous system, blood system, respiratory system and other systems. The incidence of SLE in China is about 3 times that of European and American populations, and SLE patients in China have an earlier onset of disease, more complex and diverse clinical symptoms, and are more likely to develop lupus nephritis.

[0003] At present, although great progress has been made in the diagnosis and treatment of SLE, due to the diverse initial symptoms of SLE patients and the coincidence of their clinical manifestations with other common diseases, the diagnosis usually takes a long time, and therefore it is necessary to develop new indicators for early screening to achieve the purpose of early diagnosis and treatment.

[0004] Gene expression profiling is considered a valuable tool that can provide important details for better understanding of the pathogenesis of various diseases. Peripheral blood represents the main immune cell pool of tissue / organ-infiltrating immune cells. The overall immune dysregulation commonly observed in SLE can be reflected by different changes in the composition and transcriptional state of peripheral immune cells. Traditional gene expression analysis of SLE is performed by transcriptome analysis of whole peripheral blood, which reveals the SLE type I interferon (IFN-I) signature. However, granulocytes are the most abundant circulating leukocyte type and often cause "transcriptomic noise" to mask gene expression differences in other immune cells. To avoid this problem, extensive transcriptomic analysis of SLE and RA or recent single-cell RNA sequencing (scRNA-seq) mainly focuses on peripheral blood mononuclear cells (PBMCs) or tissue-infiltrating mononuclear cells. However, this technique can largely ignore the role of neutrophils in the pathogenesis of these diseases. SUMMARY

[0005] The purpose of the present application is to provide a molecular diagnostic marker for systemic lupus erythematosus based on neutrophil signatures.

[0006] In a first aspect, the present application provides use of a substance for detecting expression amount of IFIH1 gene and / or CFLAR gene in preparation of a product for assisting in diagnosis or screening or assisting in screening of systemic lupus erythematosus.

[0007] In a second aspect, the present application provides use of a substance for detecting expression amount of IFIH1 gene and CFLAR gene and a carrier loaded with a following model formula in preparation of a product for assisting in diagnosis or screening or assisting in screening of systemic lupus erythematosus.

[0008] The model formula is:

[0009] C = 29.657 * a + 5.906 * b - 4.266,

[0010] wherein C is a value of a joint index C, a is the expression amount of IFIH1 gene, and b is the expression amount of CFLAR gene.

[0011] The expression amount of the gene is a relative expression amount.

[0012] The subject to be detected can be a suspected patient of systemic lupus erythematosus.

[0013] In the first aspect or the first aspect,

[0014] The substance for detecting expression amount of IFIH1 gene and / or CFLAR gene is any one of the following:

[0015] 1) a primer pair for amplifying IFIH1 gene and / or CFLAR gene;

[0016] 2) a PCR reagent or kit containing the primer pair.

[0017] The substance for detecting expression amount of IFIH1 gene and / or CFLAR gene in 1) above further comprises an amplification primer of a reference gene GAPDH.

[0018] The expression amount of IFIH1 gene or CFLAR gene in the above is a relative expression amount of IFIH1 or CFLAR gene, specifically, a Ct value of a target gene (IFIH1 or CFLAR) minus a Ct value of a reference gene (GAPDH) to obtain a ΔCt value; and then 2 –ΔCt is recorded as a relative expression amount of the target gene.

[0019] In a third aspect, the present application provides a kit for assisting in diagnosis of systemic lupus erythematosus, which comprises the substance for detecting expression amount of IFIH1 gene and CFLAR gene in the first or second aspect.

[0020] The kit described above further comprises a carrier loaded with the following model formula.

[0021] The model formula is C=29.657*a+5.906*b-4.266, wherein C is the value of the joint index C, a is the expression amount of the IFIH1 gene, and b is the expression amount of the CFLAR gene.

[0022] In a fourth aspect, the present application provides a device for assisting in diagnosing systemic lupus erythematosus, comprising a data acquisition module, a threshold value acquisition module, a judgment module and an output module.

[0023] The data acquisition module is configured to obtain the expression amount A of the IFIH1 gene or the CFLAR gene in the neutrophil granulocyte in the peripheral blood of a subject.

[0024] The threshold value acquisition module is configured to take the critical value at the maximum Youden index in the ROC curve of the expression amount of the IFIH1 gene or the CFLAR gene in the systemic lupus erythematosus group and the control group as the threshold value B.

[0025] The threshold value is derived from a training set, which comprises m systemic lupus erythematosus patients (in an embodiment, 46 patients) and n control subjects (in an embodiment, 55 control subjects, of which 22 are healthy and 33 are disease controls).

[0026] The judgment module is configured to compare the expression amount A with the threshold value B to obtain a judgment result, specifically as follows:

[0027] If the expression amount A is greater than the threshold value B, the subject is judged to be or is a candidate for a systemic lupus erythematosus patient.

[0028] If the expression amount A is less than or equal to the threshold value B, the subject is judged not to be or is not a candidate for a systemic lupus erythematosus patient.

[0029] The output module is configured to output the judgment result obtained by the judgment module.

[0030] In an embodiment, the expression amount A of the IFIH1 gene or the CFLAR gene in the neutrophil granulocyte in the peripheral blood is obtained by real-time fluorescent quantitative PCR detection.

[0031] In a fifth aspect, the present application provides a device for assisting in diagnosing systemic lupus erythematosus, comprising a data acquisition module, a threshold value acquisition module, a judgment module and an output module.

[0032] The data acquisition module is configured to obtain the expression amount of the IFIH1 gene and the CFLAR gene in the neutrophil granulocyte in the peripheral blood of a subject, and then calculate the value of the joint index C by a model formula; the model formula is as follows: C=29.657*a+5.906*b-4.266, wherein C is the value of the joint index C, a is the expression amount of the IFIH1 gene, and b is the expression amount of the CFLAR gene.

[0033] The threshold obtaining module is configured to draw a ROC curve of the combined indicator C value of the SLE group and the control group, and take the critical value at the maximum Youden index as the threshold D.

[0034] The control group is healthy subjects and / or disease control patients.

[0035] The threshold is derived from a training set, which includes m SLE patients (46 in an embodiment) and n control subjects (55 in an embodiment, including 22 healthy subjects and 33 disease control patients).

[0036] The judging module is configured to compare the combined indicator C value with the threshold D to obtain a judgment result, specifically as follows:

[0037] If the combined indicator C value is greater than the threshold D, the subject is judged to be or is a candidate for an SLE patient.

[0038] If the combined indicator C value is less than or equal to the threshold D, the subject is judged not to be or is not a candidate for an SLE patient.

[0039] The output module is configured to output the judgment result obtained by the judging module.

[0040] In a sixth aspect, the present application provides a product for assisting in the diagnosis of SLE, which includes the substance for detecting the expression amount of the IFIH1 gene and / or the CFLAR gene in the first or second aspect, the kit in the third aspect, or the device in the fourth or fifth aspect.

[0041] In a seventh aspect, the present application provides the use of the substance for detecting the expression amount of the IFIH1 gene and / or the CFLAR gene in the first or second aspect, the kit in the third aspect, or the device in the fourth or fifth aspect in the preparation of a product for assisting in the diagnosis of SLE.

[0042] The present application focuses on the unique changes of neutrophils in SLE, selects the neutrophil-specific gene expression markers IFIH1 and / or CFLAR, assists in the early differential diagnosis of SLE, and constructs a model for the joint diagnosis of IFIH1 and CFLAR, thereby achieving the auxiliary diagnosis or screening of SLE, and has high sensitivity and specificity. BRIEF DESCRIPTION OF DRAWINGS

[0043] Figure 1 The expression amount difference of the IFIH1 and CFLAR genes among SLE, rheumatoid arthritis, and healthy controls.

[0044] Figure 2ROC curves of the IFIH1 gene or CFLAR gene in the training set systemic lupus erythematosus group compared to the control group.

[0045] Figure 3 The results show the expression levels of the IFIH1 and CFLAR genes in the training and validation sets.

[0046] Figure 4 The left figure shows the combined C-value of the systemic lupus erythematosus group versus the control group in the training and validation sets, and the right figure shows the ROC curve of the combined C-value of the training set. Detailed Implementation

[0047] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0048] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0049] The characteristics of the case samples in the following examples are shown in Table 1:

[0050] Table 1 shows the basic characteristics of the disease group, healthy control group, and disease control group included in the study.

[0051]

[0052] Note: Systemic lupus erythematosus (SLE) refers to patients clinically diagnosed according to the diagnostic classification criteria for SLE jointly published by the European League Against Rheumatism (EULAR) and the American College of Rheumatology (ACR) in 2019; the healthy control group (HCs) included healthy volunteers with normal indicators in the physical examination center; the disease control group (DCs) included patients diagnosed according to the diagnostic criteria for various diseases, including 10 cases of rheumatoid arthritis, 10 cases of connective tissue disease, 3 cases of systemic scleroderma, 6 cases of antiphospholipid syndrome, 9 cases of Sjögren's syndrome, and 7 cases of novel coronavirus infection.

[0053] Example 1: Discovery and establishment of methods for diagnosing systemic lupus erythematosus (SLE) biomarkers IFIH1 and CFLAR genes.

[0054] I. Discovery of IFIH1 and CFLAR genes, biomarkers for diagnosing systemic lupus erythematosus.

[0055] The inventors previously analyzed the peripheral blood neutrophil transcriptomes of 14 patients with systemic lupus erythematosus, 15 patients with rheumatoid arthritis, and 13 healthy individuals (population information is shown in Table 2).

[0056] Table 2 shows the basic characteristics of systemic lupus erythematosus, rheumatoid arthritis, and healthy controls.

[0057]

[0058] *p-values ​​compare the differences between systemic lupus erythematosus (SLE) and healthy controls.

[0059] #p-value comparison between patients with systemic lupus erythematosus and rheumatoid arthritis

[0060] Differential analysis revealed significantly elevated expression of interferon-stimulated genes and cell death-related genes in patients with systemic lupus erythematosus (SLE). Using bioinformatics analysis and machine learning algorithms, the inventors screened two genes, IFIH1 and CFLAR, from among the interferon-stimulated genes and cell death-related genes. The gene expression of these genes showed significant differences among SLE patients, rheumatoid arthritis patients, and healthy controls (e.g., ...). Figure 1 (As shown).

[0061] As can be seen from the above, the expression levels of the IFIH1 gene or CFLAR gene differ significantly among healthy individuals, disease control groups, and patients with systemic lupus erythematosus (SLE), indicating that the IFIH1 gene or CFLAR gene can be used to assist in the diagnosis of SLE patients.

[0062] II. Establishment of a method for single-gene diagnosis of systemic lupus erythematosus

[0063] 1. RNA was extracted from peripheral blood neutrophils of the subjects and reverse transcribed to obtain cDNA.

[0064] Specifically as follows:

[0065] 1) Collect 2 ml of peripheral EDTA-anticoagulated blood from the subjects (including clinically diagnosed systemic lupus erythematosus patients, healthy controls, and disease controls);

[0066] 2) Add 2 ml of Ficoll / Hypaque liquid (GE Healthcare) to a 15 ml clean centrifuge tube, tilt the centrifuge tube at 45 degrees, and slowly add the anticoagulated blood sample along the tube wall with a pipette to form a clear boundary line with the Ficoll liquid. Tighten the cap and place the entire tube into a horizontal rotor centrifuge. Centrifuge at 1440 rpm (slow rise and slow fall) at room temperature for 20 minutes.

[0067] 3) Remove the centrifuge tube and carefully remove the upper plasma layer, white film layer and Ficoll separation solution in sequence using a pipette;

[0068] 4) Carefully add 600 μL of 20% polyglucose solution along the tube wall, tighten the cap, and slowly invert to mix for 5 minutes;

[0069] 5) Carefully add 4 ml of phosphate-buffered saline (PBS) along the tube wall, tighten the cap, slowly invert and mix for 5 minutes, then let stand at room temperature for 20 minutes.

[0070] 6) Once the liquid in the tube has settled to the point where a clear dividing line appears, carefully aspirate the clear liquid from the top layer into another 15 ml clean centrifuge tube using a pipette. Add 10 ml of PBS, tighten the cap, gently invert and mix, and place the entire tube into a horizontal rotor centrifuge. Centrifuge at 1500 rpm (rapid rise and fall) at room temperature for 5 minutes.

[0071] 7) Use a pipette to remove the supernatant liquid, leaving the cell pellet at the bottom of the tube. Slowly add 3 ml of ACK red blood cell lysis buffer along the tube wall, and slowly resuspend the cell pellet with a pipette. Let it stand at room temperature for 5 minutes.

[0072] 8) Carefully add 10 ml of PBS along the tube wall, tighten the tube cap, gently invert to mix, and place the whole tube into a horizontal rotor centrifuge. Centrifuge at 1500 rpm (rapid rise and fall) at room temperature for 5 minutes.

[0073] 9) Use a pipette to remove the supernatant, leaving the cell pellet at the bottom of the tube. Carefully add 700 μL of TRIzol solution along the tube wall to resuspend the cell pellet. Collect it into a 1.5 mL EP tube and let it stand at room temperature for 5 minutes.

[0074] 10) Carefully add 700 μL of anhydrous ethanol to the EP tube, tighten the cap, and invert to mix for 2 minutes;

[0075] 11) Using column extraction (Direct-zol) TM RNA was extracted using RNA MiniPrep (ZYMO RESEARCH) and placed on ice.

[0076] 12) Using the TAKARA reverse transcription kit, reverse transcribe 20 μL of the obtained mRNA into cDNA.

[0077] 2. Detection of the relative expression levels of the IFIH1 or CFLAR gene

[0078] Using the cDNA obtained in step 1 above as a template, IFIH1 and CFLAR as target genes, and GAPDH as an internal reference gene, primers were designed for real-time quantitative PCR to obtain the relative expression levels of the IFIH1 or CFLAR gene in the subjects; details are as follows:

[0079] Using PowerUp TM SYBR TMReal-time quantitative PCR experiments were performed using Green Master Mix (Thermo Fisher Scientific). IFIH1 (Forward 5'-TCACAAGTTGATGGTCCTCAAGT-3' sequence 1, Reverse 5'-CTGATGAGTTATTCTCCATGCCC-3' sequence 2), CFLAR (Forward 5'-TGCTCTTTTTGTGCCGGGAT-3' sequence 3, Reverse 5'-CGACAGACAGCTTACCTCTTTC-3' sequence 4) and GAPDH (Forward 5'-GGAGCGAGATCCCTCCAAAAT-3' sequence 5, Reverse 5'-GGCTGTTGTCATACTTCTCATGG-3' sequence 6) were used as primers, with three auxiliary wells prepared for each well, following the product instructions.

[0080] Experimental results according to 2 –ΔΔCt The analysis was performed as follows: a) the average Ct value of three parallel experiments for each gene in each sample was taken; b) the Ct value of the target gene (IFIH1 or CFLAR) in each sample was subtracted from the Ct value of the internal reference gene (GAPDH) to obtain the ΔCt value; c) the relative expression level of the target gene was then calculated as 2. –ΔCt .

[0081] 3. Determination of threshold and judgment criteria

[0082] The subjects shown in Table 1 were used as the test samples, including 71 clinically diagnosed patients with systemic lupus erythematosus (also known as test samples) and 74 control group patients (including 29 healthy controls and 45 disease controls).

[0083] All samples were randomly divided into a training set and a validation set in a 7:3 ratio. The training set included 46 patients with systemic lupus erythematosus (SLE) and 55 control patients (22 healthy individuals and 33 disease controls). The validation set included 25 patients with SLE and 19 control patients (7 healthy individuals and 12 disease controls).

[0084] The threshold is determined based on the relative expression levels of the IFIH1 and CFLAR genes in each sample of the training set. Specifically, the relative expression levels of the IFIH1 or CFLAR genes in the systemic lupus erythematosus group of the training set compared with the control group are plotted as ROC curves, and the critical value at the maximum value of the Youden index is taken as the threshold.

[0085] ROC curve as follows Figure 2 As shown, the threshold for the relative expression level of the IFIH1 gene is 0.1070; the threshold for the relative expression level of the CFLAR gene is 0.3895.

[0086] If the relative expression level of the IFIH1 gene or CFLAR gene in a subject is greater than the threshold, then the subject is or is a candidate for systemic lupus erythematosus (SLE). If the relative expression level of the IFIH1 gene or CFLAR gene in a subject is less than or equal to the threshold, then the subject is not or is not a candidate for systemic lupus erythematosus (SLE).

[0087] Example 2: Application in the diagnosis of systemic lupus erythematosus

[0088] 1. Testing

[0089] The subjects shown in Table 1 were used as the test samples, including 71 clinically diagnosed patients with systemic lupus erythematosus (also known as test samples) and 74 control group patients (including 29 healthy controls and 45 disease controls).

[0090] All samples were randomly divided into a training set and a validation set in a 7:3 ratio. The training set included 46 patients with systemic lupus erythematosus (SLE) and 55 control patients (22 healthy individuals and 33 disease controls). The validation set included 25 patients with SLE and 19 control patients (7 healthy individuals and 12 disease controls).

[0091] The detection was performed according to the method of Example 1, Part 2.

[0092] Table 1 shows the expression levels of IFIH1 and CFLAR genes in the training and validation sets of the samples. Figure 3 As shown, there are significant differences in the relative expression levels of the IFIH1 or CFLAR genes between patients with systemic lupus erythematosus and control patients in both the training and validation sets, further confirming that these two genes can serve as diagnostic biomarkers for patients with systemic lupus erythematosus.

[0093] 2. Sensitivity and Specificity Detection

[0094] ROC curves were plotted on the expression levels of IFIH1 and CFLAR genes in each systemic lupus erythematosus patient and control group in the training set. The threshold value was taken at the maximum Yoden index. Diagnostic predictions were then performed on the training set and validation set samples, and their sensitivity, specificity, positive predictive value, and negative predictive value were calculated.

[0095] The results are shown in Tables 3 and 4, which list the diagnostic sensitivity, specificity, positive predictive value, and negative predictive value, respectively. It can be seen that the expression level of IFIH1 gene or CFLAR gene alone can be used as a marker to assist in the diagnosis of patients with systemic lupus erythematosus, and the sensitivity and specificity are high.

[0096] Table 3 shows the sensitivity, specificity, positive predictive value, and negative predictive value of the IFIH1 gene.

[0097]

[0098] Table 4 shows the sensitivity, specificity, positive predictive value, and negative predictive value of the CFLAR gene.

[0099]

[0100]

[0101] Example 3: Combined diagnosis of systemic lupus erythematosus

[0102] I. Formula for Combined Diagnostic Model

[0103] As can be seen from Examples 1 and 2 above, the relative expression levels of either the IFIH1 gene or the CFLAR gene alone can be used to assist in the diagnosis of systemic lupus erythematosus (SLE) patients. To further investigate whether the combined diagnosis of the two can improve diagnostic efficacy, a combined diagnostic model formula was constructed based on the training set results:

[0104] C = 29.657*a + 5.906*b - 4.266

[0105] Where C is the combined index C value, a is the relative expression level of the IFIH1 gene, and b is the relative expression level of the CFLAR gene.

[0106] The relative expression levels of the aforementioned IFIH1 or CFLAR genes are 2. –ΔCt The ΔCt value is the Ct value of the target gene (IFIH1 or CFLAR) minus the Ct value of the internal reference gene (GAPDH).

[0107] II. Establishment of a method for dual-gene diagnosis of systemic lupus erythematosus

[0108] 1. Same as step 1 in Example 1, cDNA is obtained;

[0109] 2. Same as step 2 in Example 1, the relative expression levels of the IFIH1 gene and the CFLAR gene were obtained.

[0110] 3. Substitute the relative expression levels of the target gene IFIH1 and the target gene CFLAR obtained in step 2 above into the following combined diagnostic model formula to calculate the combined index C value.

[0111] C = 29.657*a + 5.906*b - 4.266

[0112] Where C is the combined index C value, a is the relative expression level of the IFIH1 gene, and b is the relative expression level of the CFLAR gene.

[0113] ROC curves were plotted based on the combined C-value of the systemic lupus erythematosus group versus the control group in the training set. Figure 4 (See the right figure). Taking the critical value at the maximum value of the Youden index, the threshold for obtaining the joint index C value is 0.1535.

[0114] If a subject's combined indicator C value is greater than the threshold, then the subject is or is a candidate for systemic lupus erythematosus (SLE). If a subject's combined indicator C value is less than or equal to the threshold, then the subject is not or is not a candidate for systemic lupus erythematosus (SLE).

[0115] III. Application of dual-gene diagnosis in systemic lupus erythematosus

[0116] The subjects shown in Table 1 were used as the test samples, including 71 clinically diagnosed patients with systemic lupus erythematosus (also known as test samples) and 74 control group patients (including 29 healthy controls and 45 disease controls).

[0117] All samples were randomly divided into a training set and a validation set in a 7:3 ratio. The training set included 46 patients with systemic lupus erythematosus (SLE) and 55 control patients (22 healthy individuals and 33 disease controls). The validation set included 25 patients with SLE and 19 control patients (7 healthy individuals and 12 disease controls).

[0118] Following the method described in section two above, the relative expression levels of the IFIH1 and CFLAR genes from each systemic lupus erythematosus patient and control group in the training and validation sets are substituted into the aforementioned joint diagnostic model formula to calculate the C-value for each subject. Figure 4 (Left image).

[0119] ROC curves were plotted using the C-values ​​of each systemic lupus erythematosus patient and control group in the training set, and the threshold value was taken as the value at the point where the maximum Yoden index was reached. Figure 4 (See the right figure). Diagnostic predictions are performed on the training set and validation set samples respectively, and their sensitivity, specificity, positive predictive value and negative predictive value are calculated.

[0120] The results are shown in Table 5. It can be seen that the combined expression levels of IFIH1 and CFLAR genes can be used to assist in the diagnosis of systemic lupus erythematosus patients, with high sensitivity and specificity.

[0121] Table 5 shows the sensitivity, specificity, positive predictive value, and negative predictive value of the combined diagnostic C-value.

[0122]

Claims

1. Detection of peripheral blood neutrophils IFIH1 Genes and CFLAR The application of substances that enhance gene expression in the preparation of products for the auxiliary diagnosis or screening of systemic lupus erythematosus.

2. Detection of peripheral blood neutrophils IFIH1 Genes and CFLAR The application of the gene expression level material and the vector with the following model formula in the preparation of products for auxiliary diagnosis or screening or auxiliary screening of systemic lupus erythematosus; The model formula is as follows: C = 29.657 * a + 5.906 * b - 4.266, Where C is the joint index C value, and a is... IFIH1 Gene expression level, b is CFLAR Gene expression levels.

3. The application according to claim 1 or 2, characterized in that: The detection IFIH1 Genes and CFLAR The substance that determines gene expression is any of the following: 1) Includes amplification IFIH1 Genes and CFLAR Primer pairs for genes; 2) PCR reagents or kits containing the primer pairs described above.

4. A device for assisting in the diagnosis of systemic lupus erythematosus, comprising a data acquisition module, a threshold acquisition module, a judgment module, and an output module; The data acquisition module is used to obtain data from the peripheral blood neutrophils of the subject. IFIH1 Genes and CFLAR Gene expression level A; The threshold acquisition module is used to obtain the threshold values ​​from the systemic lupus erythematosus group compared to the control group. IFIH1 Genes and CFLAR The critical value at the maximum value of the Youden exponent in the ROC curve of gene expression is used as the threshold B; The judgment module is used to compare the expression level A with the threshold B to obtain a judgment result. The output module is used to output the judgment result obtained by the judgment module.

5. A device for assisting in the diagnosis of systemic lupus erythematosus, comprising a data acquisition module, a threshold acquisition module, a judgment module, and an output module; The data acquisition module is used to obtain data from the peripheral blood neutrophils of the subject. IFIH1 Genes and CFLAR The gene expression level is then used to calculate the combined index C value using a model formula; the model formula is as follows: C = 29.657 * a + 5.906 * b - 4.266, where C is the combined index C value, and a is... IFIH1 Gene expression level, b is CFLAR Gene expression levels; The threshold acquisition module is used to plot the combined index C value of the systemic lupus erythematosus group and the control group on an ROC curve, and take the critical value at the maximum value of the Youden index as the threshold D. The judgment module is used to compare the joint index C value with the threshold D to obtain a judgment result. The output module is used to output the judgment result obtained by the judgment module.

6. The detection method according to any one of claims 1-3 IFIH1 Genes and CFLAR Application of substances that enhance gene expression in the preparation of products for the auxiliary diagnosis of systemic lupus erythematosus.

7. The application of the following kit in the preparation of products for the auxiliary diagnosis of systemic lupus erythematosus; The kit contains the detection method for peripheral blood neutrophils as described in claim 2. IFIH1 Genes and CFLAR The substance for gene expression and the vector for the loading model formula.

8. The use of the device according to claim 4 or 5 in the preparation of products for the auxiliary diagnosis of systemic lupus erythematosus.

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