Blau syndrome biomarkers and their applications

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to a Blau syndrome biomarker and its application. The biomarker is selected from IL-8 and / or SDF-1. The biomarker has a high AUC value, sensitivity and specificity. By detecting the expression of IL-8 and / or SDF-1, it is possible to distinguish Blau syndrome from other pediatric inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and pediatric idiopathic uveitis (PIU)) quickly, accurately, simply, safely and at low cost, thereby improving the accuracy of Blau syndrome diagnosis.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to a Blau syndrome biomarker and application thereof. Background Art

[0002] Blau syndrome (BS), also known as childhood granulomatous arthritis, is an early-onset autoinflammatory disease with an estimated incidence of 0.05 cases per 100,000 person-years. It is inherited in an autosomal dominant manner and is widely believed to be caused by mutations in the NOD2 gene (such as one or more mutations in R334W, R334Q, E383K, G481D, W490S, M513T, R587C, and N670K). Its typical features are the triad of granulomatous dermatitis, symmetrical arthritis, and recurrent uveitis. In addition to specific manifestations (such as fever), it may also be accompanied by multi-system involvement, such as large-vessel vasculitis, sialadenitis, lymphadenopathy, granulomatous tubular and interstitial nephritis, and granulomatous changes in the liver and spleen. Blau syndrome develops in early childhood and may cause joint degeneration. 60%-80% of Blau syndrome cases may also involve the eyes, and some other eye diseases (such as chorioretinitis, cataracts, glaucoma, and retinal detachment) may complicate eye involvement, leading to severe visual impairment or even blindness. Therefore, the diagnosis of Blau syndrome is particularly important.

[0003] However, due to the rarity of Blau syndrome and the overlap of its clinical symptoms with other pediatric inflammatory diseases, especially juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU), the disease is prone to misdiagnosis and missed diagnosis. In addition, the development of the disease is more serious than that of JIA and PIU. Misdiagnosis and missed diagnosis may lead to insufficient treatment and affect long-term prognosis. For example, clinical analysis shows that 45.2% of Blau syndrome patients develop secondary glaucoma, compared with only 16.7% of JIA and 21.3% of PIU patients. For another example, Blau syndrome patients also have a higher incidence of rash (50.0%), joint deformity (64.5%) and small joint deformity (43.5%), while the incidence of rash (5.6% and 3.9%, respectively) and joint deformity (25.0% and 0.2%) in JIA and PIU patients are significantly lower than those in Blau syndrome patients. Therefore, early identification of Blau syndrome can achieve timely treatment, prevent or delay the occurrence of severe symptoms in patients, and reduce long-term disability.

[0004] Currently, the diagnosis of Blau syndrome mainly includes skin and / or synovial pathological biopsy and genetic testing. However, skin and / or synovial pathological biopsy has the following disadvantages: (1) Pathological biopsy is an invasive operation, which may be accompanied by a certain risk of complications such as bleeding and infection; (2) The quality of the biopsy sample has an important impact on the final result. If the quality of the collected sample is poor, it may affect the accuracy and reliability of the pathological examination and increase the difficulty of detection; (3) Skin tissue biopsy can only provide histological information of the local area and cannot fully reflect the situation of systemic diseases. It may require biopsy of multiple sites or combined with other examination methods to obtain more comprehensive information, and the operation is cumbersome; (4) The skin and synovial pathological characteristics of Blau syndrome are similar to other inflammatory joint lesions. This non-specific manifestation may lead to difficulties in diagnosis and requires comprehensive analysis in combination with other clinical and laboratory tests. The operation is also cumbersome. In addition, genetic testing also has some disadvantages: (1) Genetic testing is costly, has complex operating procedures, and is highly dependent on technical personnel, which limits its application in routine clinical practice; (2) The sequencing technology of genetic testing takes a long time and cannot be used for rapid on-site testing, which affects the patient's diagnosis and treatment process. Summary of the invention

[0005] In response to the above problems, the present invention has discovered biomarkers IL-8 and / or SDF-1 associated with Blau syndrome, both of which have high AUC values, sensitivity and specificity. Based on clinical symptoms, by detecting the expression levels of IL-8 and / or SDF-1, Blau syndrome can be distinguished from other childhood inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU)) in a rapid, accurate, simple, safe and low-cost manner, thereby achieving the purpose of accurate prognosis of Blau syndrome.

[0006] In order to achieve the above object, the present invention can adopt the following technical solutions:

[0007] In one aspect, the present invention provides a Blau syndrome biomarker, wherein the biomarker is selected from IL-8 and / or SDF-1.

[0008] Another aspect of the present invention provides use of a reagent for detecting a Blau syndrome biomarker in preparing a product for diagnosing Blau syndrome, wherein the Blau syndrome biomarker is selected from IL-8 and / or SDF-1.

[0009] The beneficial effects of the present invention include at least:

[0010] (1) ROC curve analysis of the Blau syndrome biomarker IL-8 provided by the present invention shows that its AUC can reach 0.772, the sensitivity value can reach 0.611, and the specificity value can reach 0.824, that is, it has good AUC, sensitivity and specificity. By detecting the IL-8 expression level combined with clinical symptoms, Blau syndrome can be accurately distinguished from other childhood inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU)).

[0011] (2) ROC curve analysis of the Blau syndrome biomarker SDF-1 provided by the present invention shows that its AUC can reach 0.624, the sensitivity value can reach 0.778, and the specificity value can reach 0.548, that is, it has good AUC, sensitivity and specificity. By detecting the SDF-1 expression level combined with clinical symptoms, Blau syndrome can be accurately distinguished from other childhood inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU)).

[0012] (3) The Blau syndrome biomarkers SDF-1 and IL-8 (combined) provided by the present invention, ROC curve analysis shows that their AUC can reach 0.820, the sensitivity value can reach 0.722, and the specificity value can reach 0.857, that is, they have good AUC, sensitivity and specificity. By detecting the expression levels of SDF-1 and IL-8 combined with clinical symptoms, Blau syndrome can be accurately distinguished from other childhood inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU)). BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The significantly different expression of IL-8 in aqueous humor samples in the embodiments of the present invention;

[0014] Figure 2 is the significantly differential expression of SDF-1 in aqueous humor samples in the embodiment of the present invention; Mut represents Blau syndrome patients with classic NOD2 mutations (such as one or more mutations among R334W, R334Q, E383K, G481D, W490S, M513T, R587C and N670K), wherein Nomut represents pediatric uveitis patients without NOD2 mutations (including uveitis associated with juvenile idiopathic arthritis and pediatric idiopathic uveitis), and Ctrl represents patients with senile cataracts;

[0015] Figure 3 Shown are the ROC curves for the combined diagnosis of SDF-1, IL-8, and SDF-1+IL-8 in aqueous humor samples. DETAILED DESCRIPTION

[0016] The examples are provided to better illustrate the present invention, but the present invention is not limited to the examples. Therefore, those skilled in the art may make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still fall within the protection scope of the present invention.

[0017] The terms used herein are only used to describe specific embodiments and are not intended to limit the present disclosure. Unless the context has a significantly different meaning, expressions in the singular include expressions in the plural. As used herein, it should be understood that terms such as "include", "have", "include" are intended to indicate the presence of features, numbers, operations, components, parts, elements, materials or combinations. The terms of the present invention are disclosed in the specification, and are not intended to exclude the possibility that one or more other features, numbers, operations, components, parts, elements, materials or combinations thereof may exist or may be added. As used herein, " / " may be interpreted as "and" or "or", depending on the circumstances.

[0018] An embodiment of the present invention provides a Blau syndrome biomarker, wherein the biomarker is selected from IL-8 and / or SDF-1.

[0019] It should be noted that SDF-1 (stromal cell-derived factor 1) and IL-8 (interleukin-8) are two cytokines related to immune response and inflammation. Among them, SDF-1, also known as CXCL12, is a chemokine that plays a key role in inflammation and tissue repair. It binds to the CXCR4 receptor and guides immune cells to migrate to damaged tissues, participating in inflammatory responses and the localization of immune cells; IL-8 is a proinflammatory cytokine, also known as chemokine CX-Cmotif ligand 8 (CXCL8), which is mainly produced by a variety of cell types, including macrophages, epithelial cells, and endothelial cells; IL-8 binds to CXCR1 and CXCR2 receptors, attracting neutrophils and other inflammatory cells to migrate to the site of inflammation, participating in the regulation of inflammatory responses. The present invention found that the expression levels of IL-8 and SDF-1 in the aqueous humor of Blau syndrome patients with classic NOD2 mutations (such as one or more mutations among R334W, R334Q, E383K, G481D, W490S, M513T, R587C and N670K) were significantly different from those in children with uveitis (including patients with juvenile idiopathic arthritis and children with idiopathic uveitis) without NOD2 mutations (specifically, the expression level of IL-8 was significantly increased compared with that in children with uveitis without NOD2 mutations, and the expression level of SDF-1 was significantly increased compared with that in children with uveitis without NOD2 mutations). The ROC curve showed that IL-8 and SDF-1 as well as IL-8 combined with SDF-1 had higher AUC values, sensitivity values ​​and specificity values, so IL-8 and SDF-1 can be used alone as markers for the diagnosis of Blau syndrome, and IL-8 and SDF-1 combined can also be used as markers for the diagnosis of Blau syndrome; that is, Blau syndrome can be distinguished from other childhood inflammatory diseases (such as juvenile idiopathic arthritis (JIA) and childhood idiopathic uveitis (PIU)) based on the expression level of IL-8 and / or SDF-1 combined with clinical symptoms.

[0020] The embodiment of the present invention also provides a use of a reagent for detecting a Blau syndrome biomarker in preparing a product for diagnosing Blau syndrome, wherein the Blau syndrome biomarker is selected from IL-8 and / or SDF-1.

[0021] It should be noted that, as mentioned above, IL-8 and SDF-1 alone or in combination can be used as markers for diagnosing Blau syndrome; therefore, Blau syndrome can be diagnosed by using reagents for detecting IL-8 or SDF-1 or reagents for detecting IL-8 or SDF-1; it should be understood that the reagents for detecting Blau syndrome biomarkers in the present invention include the following situations: (a) reagents that can detect IL-8; (b) reagents that can detect SDF-1; (c) reagents that can simultaneously detect IL-8 and SDF-1; (d) reagents that can detect IL-8 and reagents that can detect SDF-1, and the reagents that can detect IL-8 and the reagents that can detect SDF-1 are different.

[0022] It should be noted that the use of reagents based on the detection of Blau syndrome biomarkers in the diagnosis of Blau syndrome has the following advantages over traditional diagnostic methods: (1) Sensitivity and specificity: Compared with pediatric uveitis (juvenile idiopathic arthritis and pediatric idiopathic uveitis) with similar clinical symptoms, IL-8 and SDF-1 in the body fluid samples of Blau syndrome patients are significantly increased, which can more accurately diagnose the disease with high sensitivity and specificity, providing clinicians with new targets for accurate diagnosis; (2) Rapidity: Compared with tissue biopsy or genetic testing, the detection of IL-8 and SDF-1 usually has a shorter detection cycle, which can obtain diagnostic results more quickly and help to take timely treatment measures; (3) Repeatability: The cytokine detection method has good repeatability and stability, and can be tested multiple times at different time points, which helps to track the progression of the disease and the effect of treatment.

[0023] In some specific embodiments, the reagents for detecting Blau syndrome biomarkers include one or more of the following reagents: (1) reagents for detecting the expression of IL-8 and / or SDF-1 by immunodetection methods; (2) reagents for detecting the expression of IL-8 and / or SDF-1 by flow cytometry; (3) reagents for detecting the expression of IL-8 and / or SDF-1 by immunohistochemistry; (4) reagents for detecting the expression of IL-8 and / or SDF-1 by PCR amplification or real-time quantitative PCR amplification; (5) reagents for detecting the expression of IL-8 and / or SDF-1 by immunoblotting; (6) reagents for detecting the expression of IL-8 and / or SDF-1 by RNA-seq.

[0024] It should be noted that both IL-8 and SDF-1 are proteins, so the above-mentioned reagent for detecting Blau syndrome biomarkers can be a reagent for detecting the expression level of IL-8 and / or SDF-1 by the above-mentioned detection method.

[0025] In some specific embodiments, the reagents for detecting the expression of IL-8 and / or SDF-1 by immunodetection methods include one or more of the following reagents: (1) antibodies that specifically bind to IL-8; (2) antibodies that specifically bind to SDF-1; (3) antibodies that bind to both IL-8 and SDF-1; (4) antibodies that specifically bind to IL-8 and antibodies that specifically bind to SDF-1.

[0026] It should be noted that, as described above, the reagent for detecting the Blau syndrome biomarker in the present invention can be a reagent for detecting the expression of IL-8 and / or SDF-1 by an immunoassay method, especially an antibody that can specifically bind to IL-8 and / or SDF-1.

[0027] In some specific embodiments, the reagents for detecting the expression amount of IL-8 and / or SDF-1 by the immunodetection method include an enzyme-linked immunosorbent assay system or an immunofluorescence analysis system, and the enzyme-linked immunosorbent assay system also includes one or more of a blocking solution, an antibody diluent, a washing buffer, and a color stop solution; the immunofluorescence analysis system also includes one or more of a blocking solution, an antibody diluent, a washing buffer, a color stop solution, biotin, magnetic beads or a fluorescent reagent.

[0028] It should be noted that, based on the above antibodies that specifically bind to IL-8 and / or SDF-1, the expression of IL-8 and / or SDF-1 can be measured using an enzyme-linked immunosorbent assay (ELISA), and the reagent for detecting the Blau syndrome biomarker can be an enzyme-linked immunosorbent assay system (ELISA reagent) comprising one or more of a blocking solution, an antibody diluent, a washing buffer, and a color stop solution. In addition, on this basis, fluorescent reagents can be used to label antibodies for immunofluorescence analysis, and magnetic beads can be added to facilitate the separation and detection of antibodies.

[0029] In some specific embodiments, the above-mentioned reagents for detecting the expression level of IL-8 and / or SDF-1 by PCR amplification or real-time quantitative PCR amplification include one or more of the following: (1) primer pairs for amplifying IL-8 and / or SDF-1 expression genes; (2) primer pairs and probes for amplifying IL-8 and / or SDF-1 expression genes; (3) primer pairs and fluorescent dyes for amplifying IL-8 and / or SDF-1 expression genes.

[0030] It should be noted that, in addition to the above-mentioned immunoassay method, PCR amplification detection is also one of the main methods for protein detection. Amplification primers can be designed according to the expression genes of IL-8 and / or SDF-1 to detect the expression levels of IL-8 and / or SDF-1 after amplification; and, on this basis, probes or fluorescent dyes of IL-8 and / or SDF-1 can be added to realize the visual detection of IL-8 and / or SDF-1 expression.

[0031] In some specific embodiments, the reagent for detecting the expression of IL-8 and / or SDF-1 by PCR amplification or real-time quantitative PCR amplification may further include one or more of PCR enzyme, PCR buffer or dNTPS.

[0032] It should be noted that when performing PCR amplification or real-time quantitative PCR amplification, in addition to the above-mentioned primer pairs, probes or fluorescent dyes, one or more of PCR enzymes, PCR buffers or dNTPS are also included. In addition, the above-mentioned primer pairs and probes can be designed according to common knowledge, and fluorescent dyes, PCR enzymes, PCR buffers or dNTPS are all well known in the art.

[0033] In some specific embodiments, the product for diagnosing Blau syndrome is a diagnostic reagent for diagnosing Blau syndrome, a diagnostic kit for diagnosing Blau syndrome, or a diagnostic system for diagnosing Blau syndrome.

[0034] It should be noted that the above-mentioned products for diagnosing Blau syndrome can be in many forms, generally in the form of reagents and kits, which have low preparation costs, are easy to carry, and are not difficult to operate. In addition, a diagnostic system can be built based on reagents and kits, which is more conducive to patient information management and patient condition monitoring.

[0035] In some specific embodiments, the above-mentioned diagnostic system for diagnosing Blau syndrome may include the following devices: (1) an information collection device for obtaining the expression level of the biomarkers IL-8 and / or SDF-1; (2) a calculation device for calculating whether the subject suffers from Blau syndrome based on the information collected in (1); (3) a result output device for whether the subject suffers from Blau syndrome.

[0036] It should also be noted that, in the present invention, by detecting the expression of SDF-1 or IL-8 in combination with clinical symptoms, Blau syndrome can be accurately distinguished from other inflammatory diseases of children (such as juvenile idiopathic arthritis (JIA) and idiopathic uveitis (PIU) in children), thereby improving the accuracy of diagnosis of Blau syndrome; by detecting the expression of SDF-1 and IL-8 in combination with clinical symptoms, Blau syndrome can be accurately distinguished from other inflammatory diseases of children (such as juvenile idiopathic arthritis (JIA) and idiopathic uveitis (PIU) in children), thereby improving the accuracy of diagnosis of Blau syndrome. In addition, when two expression levels are detected at the same time, if only one of the expression levels is significantly different from other inflammatory diseases of children, it indicates the possibility of suffering from Blau syndrome.

[0037] In order to better understand the present invention, the content of the present invention is further explained below in conjunction with specific examples, but the content of the present invention is not limited to the following examples.

[0038] In the present invention, the human cytokine screening 48-Plex kit (purchased from Bio-rad, catalog number: 12007283) was used to strictly follow the manufacturer's instructions to detect 18 patients with Blau syndrome with typical NOD2 mutations and 33 children with uveitis without NOD2 mutations (including 15 patients with juvenile idiopathic arthritis (JIA) and 18 patients with pediatric idiopathic uveitis (PIU)) (NOD2 mutations have been verified by sequencing) and 18 patients with senile leukoderma. The aqueous humor of patients with Blau syndrome was used as a control. It was found that the expression levels of IL-8 and SDF-1 in patients with Blau syndrome with typical NOD2 mutations were significantly different from those in children with uveitis without NOD2 mutations (specifically, the expression level of IL-8 was significantly increased compared with that in children with uveitis without NOD2 mutations, and the expression level of SDF-1 was significantly decreased compared with that in children with uveitis without NOD2 mutations), and there were statistical differences (compared using analysis of variance (ANOVA), with the concentration unit in pg / mL). The specific results are as follows Figure 1 and Figure 2 shown.

[0039] In addition, two independent sample t-tests were performed on the expression levels of IL-8 and SDF-1 in the aqueous humor of 15 JIA patients and 18 Blau syndrome patients, and 18 PIU patients and 18 Blau syndrome patients. The test results are shown in Tables 1 to 9 below.

[0040] Table 1 t-test results of IL-8 expression levels in Blau syndrome and JIA

[0041]

[0042]

[0043] As can be seen from Table 1 above, the t-test significance result is 0.018, indicating that there is a significant difference in IL-8 expression levels between Blau syndrome and JIA.

[0044] Table 2 t-test results of IL-8 expression level on Blau syndrome and PIU

[0045]

[0046] As can be seen from Table 2 above, the t-test significance result is 0.028, indicating that there is a significant difference in the expression level of IL-8 between Blau syndrome and PIU.

[0047] Table 3 t-test results of SDF-1 expression level in Blau syndrome and PIU

[0048]

[0049] As can be seen from Table 3 above, the t-test significance result is 0.028, indicating that there are significant differences in SDF-1 between Blau syndrome and JIA.

[0050] Table 4 t-test results of SDF-1 expression level in Blau syndrome and PIU

[0051]

[0052] As can be seen from Table 3 above, the t-test significance result is 0.019, indicating that there is a significant difference in the expression level of SDF-1 in Blau syndrome and PIU.

[0053] Further, ROC curves were drawn to show the distinction between cases and controls, where cases refer to Blau syndrome patients with classic NOD2 mutations (one or more mutations among R334Q, R334, WE383K, W490S, R587C and G481D), while the control group includes pediatric uveitis patients without NOD2 mutations (including uveitis associated with juvenile idiopathic arthritis and pediatric idiopathic uveitis) and senile cataract patients (as controls), specifically including (in addition, the specific approach can also refer to the SPSS Statistical Analysis Basic Tutorial (author: Jiang Qianjin, Li Chuanjiang)):

[0054] (1) Start SPSS (IBM SPSS Statistics 25.0): Open SPSS25.0 software and load the data set;

[0055] (2) Data preparation: The data included 48 cytokine measurements from 18 children with Blau syndrome who had been confirmed to have NOD2 mutations by gene sequencing and 33 children with uveitis who did not have NOD2 mutations. The binary dependent variable was defined as state = 1 (positive) for children with Blau syndrome who had NOD2 mutations and state = 0 (negative) for children with uveitis who did not have NOD2 mutations.

[0056] (3) Open the ROC curve function: In the menu bar, select Analyze-ROC Curve;

[0057] (4) Select variables: In the pop-up dialog box, drag the binary dependent variable (i.e., the state assigned in step 2) to the State Variable box; drag the predictor variable (i.e., the cytokine measurement value) to the TestVariable(s) box;

[0058] (5) Define the value of the state variable: Click Define State and define it as 1;

[0059] (6) Select curve options: In the dialog box, select Display ROC Curve to draw the curve; you can select more options by clicking the Options button, such as displaying confidence intervals, selecting the positive and negative class ratios, etc.

[0060] (7) Run the analysis: After confirming that all settings are correct, click the OK button and SPSS will generate the ROC curve;

[0061] (8) View results: The analysis results will be displayed in the output window, and you can see the ROC curve, area under the curve (AUC), sensitivity and specificity table, as shown in Table 5 below.

[0062] Table 5 AUC values, 95% CI values, sensitivity and specificity of IL-8 and SDF-1

[0063]

[0064] In summary, the present invention found that the expression levels of IL-8 and SDF-1 in patients with Blau syndrome were significantly upregulated compared with those in patients with juvenile idiopathic arthritis, childhood idiopathic uveitis and controls, and ROC curve analysis showed that IL-8 and SDF-1, as well as IL-8 and SDF-1 combined as combined diagnostic factors, can be used to diagnose Blau syndrome.

[0065] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be covered by the scope of the claims of the present invention.

Claims

1. Use of a reagent for detecting a Blau syndrome biomarker in the preparation of a product for diagnosing Blau syndrome, wherein the Blau syndrome biomarker is a combination of IL-8 and SDF-1.

2. The use according to claim 1, characterized in that: Reagents for detecting Blau syndrome biomarkers include one or more of the following reagents: (1) reagents for detecting the expression of IL-8 and SDF-1 by immunoassay; (2) reagents for detecting the expression of IL-8 and SDF-1 by flow cytometry; (3) reagents for detecting the expression of IL-8 and SDF-1 by immunohistochemistry; (4) reagents for detecting the expression of IL-8 and SDF-1 by PCR amplification or real-time quantitative PCR amplification; (5) reagents for detecting the expression of IL-8 and SDF-1 by immunoblotting; and (6) reagents for detecting the expression of IL-8 and SDF-1 by RNA-seq.

3. The use according to claim 2, characterized in that: The reagents for detecting the expression levels of IL-8 and SDF-1 by immunoassay methods include one or more of the following: (1) antibodies that bind to both IL-8 and SDF-1; (2) antibodies that specifically bind to IL-8 and antibodies that specifically bind to SDF-1.

4. The use according to claim 3, characterized in that: Reagents for detecting the expression levels of IL-8 and SDF-1 by immunoassay methods include enzyme-linked immunosorbent assay system or immunofluorescence analysis. The enzyme-linked immunosorbent assay system includes one or more of a blocking solution, an antibody diluent, a washing buffer, and a color stop solution; the immunofluorescence analysis includes one or more of a blocking solution, an antibody diluent, a washing buffer, a color stop solution, biotin, magnetic beads or a fluorescent reagent.

5. The use according to claim 2, characterized in that: The reagents for detecting the expression levels of IL-8 and SDF-1 by PCR amplification or real-time quantitative PCR amplification include one or more of the following: (1) a primer pair for amplifying IL-8 and SDF-1 expression genes; (2) a primer pair and a probe for amplifying IL-8 and SDF-1 expression genes; (3) a primer pair and a fluorescent dye for amplifying IL-8 and SDF-1 expression genes.

6. The use according to claim 5, characterized in that: The reagents for detecting the expression levels of IL-8 and SDF-1 by PCR amplification or real-time quantitative PCR amplification also include one or more of PCR enzyme, PCR buffer or dNTPS.

7. The use according to any one of claims 1 to 6, characterized in that: The product for diagnosing Blau syndrome is a diagnostic reagent for diagnosing Blau syndrome or a diagnostic system for diagnosing Blau syndrome; the diagnostic system for diagnosing Blau syndrome includes the following devices: (1) an information collection device for obtaining the expression levels of biomarkers IL-8 and SDF-1; (2) a calculation device for calculating whether the subject suffers from Blau syndrome based on the information collected in (1); and (3) a result output device for whether the subject suffers from Blau syndrome.

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