Inflammation factor detection kit based on liquid-based chip technology

Abstract

The invention relates to the technical field of biological detection, and provides an inflammatory factor detection kit based on a liquid-based chip technology, and the inflammatory factor detection kit comprises six fluorescent microspheres coupled with capture antibodies, and six detection antibodies labeled by report fluorescence; in the six fluorescent microspheres containing the captured antibodies, the fluorescence intensities of all the fluorescent microspheres are different; according to the detection kit provided by the invention, on the basis of a liquid-based chip technology, fluorescein is adopted to carry out fluorescence coding on microspheres, and a captured antibody is connected to the fluorescent microspheres through a chemical reaction; a detection target is captured through the fluorescent microspheres, and then a signal is displayed through the detection antibody, so that the detection of the inflammatory factors is completed, the defects of large demand quantity of soluble inflammatory factor detection samples, long detection time, tedious detection steps and the like in the prior art are overcome, and great convenience is brought to scientific researchers at a hospital end and an industrial end.

Description

Technical Field

[0001] The present invention relates to the field of biological detection technology, and in particular to an inflammatory factor detection kit based on liquid-based chip technology. Background Art

[0002] Inflammation is a self-protective mechanism of organisms to eliminate harmful stimuli and promote healing, and its process involves complex biochemical reactions. Inflammatory reactions can be triggered by a variety of factors, mainly including the following categories: 1. Biological factors: After bacteria, viruses, fungi, parasites and other pathogens infect the human body, they can activate the immune system to trigger an inflammatory response; 2. Physical factors: Physical factors such as high temperature, low temperature, ultraviolet rays, radiation, mechanical damage, etc. can directly damage tissue cells, thereby triggering inflammation; 3. Chemical factors: Exogenous chemical substances such as strong acids, strong alkalis, corrosive substances, and endogenous toxic substances such as tissue necrosis decomposition products can directly stimulate tissues to cause inflammation; 4. Immune response: When the body produces an abnormal immune response to its own tissue antigens or foreign antigens, it can lead to tissue damage and inflammatory reactions. When the body has an inflammatory response, it will cause tissue damage, vasodilation and increased vascular permeability, and white blood cells will infiltrate and migrate to participate in the regulation of inflammatory reactions; immune cells express inflammatory factors to regulate the body's immune response, thereby removing damaging factors and repairing damaged tissues.

[0003] Inflammatory factors are a class of biologically active molecules involved in inflammatory responses, mainly produced by immune cells, endothelial cells, fibroblasts, etc. They play a key role in the initiation, development and resolution of inflammation. Among them, some inflammatory factors play a vital role in the diagnosis of some diseases.

[0004] Interleukin-2 receptor α (CD25) is a 55kDa type I transmembrane protein. It is mainly expressed in tissues such as the spleen and lymph nodes, and on the surface of Treg cells, basophils, activated T cells, etc. sCD25 is the shedding of the membrane-bound CD25 chain. sCD25 plays an important role in the diagnosis of hemophagocytic syndrome and some autoimmune diseases. 1. Soluble CD25 is of great significance in the diagnosis and prognosis of hemophagocytic syndrome (HLH). According to the research results of the domestic collaborative group and the Mayo Clinic, sCD25 levels ≥6400pg / ml can be used as one of the diagnostic indicators (HLH-2004 diagnostic criteria). sCD25 is closely related to the immediate state of severity of hemophagocytic syndrome. It usually rises before clinical deterioration and drops rapidly during the recovery of inflammatory response. The elevated level of sCD25 can help determine the prognosis and differentiate hemophagocytic syndrome from other diseases such as sepsis. 2. In novel coronavirus and active Mycobacterium tuberculosis infection, the degree of sCD25 elevation is positively correlated with the severity and prognosis of the disease. 3. sCD25 reflects the inflammatory process related to the formation of atherosclerosis, and higher levels are associated with an increased risk of myocardial infarction. In addition, sCD25 can be elevated in diseases such as multiple sclerosis and lymphogranuloma.

[0005] CD40L, also known as gp39, is a 33kD type II transmembrane glycoprotein with a total of 266aa, including 214 amino acids in the extracellular region; it is lowly expressed in activated CD4+T lymphocytes, partially activated CD8+T cells, basophils, mast cells, and NK cells. It participates in the body's humoral and cellular immune responses, and can induce the expression of a series of adhesion molecules such as intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, and E-selectin, promote the adhesion of leukocytes to blood vessels, induce immune responses, and promote the formation of atherosclerotic plaques. sCD40L is a soluble protein of the extracellular segment of the CD40L receptor protein. sCD40L in the human body fluid circulation mainly comes from activated platelets (95%) and lymphocytes. It has the dual characteristics of pro-thrombosis and pro-inflammatory, and is a marker of immune system activation. Studies have shown that sCD40L can mediate the inflammatory response in the process of atherosclerosis and can be used as a diagnostic marker for systemic atherosclerosis.

[0006] Interleukin-6 receptor β subunit, CD130, also known as gp130, is a glycoprotein with a molecular weight of 130kD and is widely expressed on a variety of mammalian cells. CD130 can produce soluble CD130 (sCD130, sgp130) through proteolysis or selective splicing. As a negative regulator, sCD130 mediates IL-6 / IL-6R signaling and exhibits anti-inflammatory properties mainly through endogenous inhibition of IL-6 trans-signaling. It participates in immune response, hematopoiesis, inflammation, and endocrine and nervous system functions. Studies have shown that the serum sCD130 concentration in patients with stable coronary atherosclerosis (CAD) is negatively correlated with the severity of coronary artery damage.

[0007] CD354, also known as myeloid cell triggering receptor 1 (TREM 1), is an immunoglobulin superfamily receptor named for its specific expression on neutrophils and monocytes, i.e. myeloid cells. TREM-1 has two forms: cell membrane (mTREM-1) and soluble (sTREM-1). Among them, sTREM-1 is the extracellular region fragment of soluble TREM-1, which plays an important role in the inflammatory stimulation and cascade amplification effect in infectious diseases. TREM-1 is specifically expressed on the surface of neutrophils and monocytes. When the body is stimulated by various pathogens, such as bacteria and viruses, its expression level increases significantly and is released into the blood or body fluids, with early onset and short half-life. Increased sTREM-1 is commonly seen in bacterial meningitis, bacterial pleural effusion, COPD combined infection and sepsis, etc. It is rarely or not expressed in non-infectious inflammatory diseases, suggesting that sTREM-1 can be used as a specific indicator for diagnosing bacterial infection.

[0008] Pentraxin 3 (PTX3), also known as tumor necrosis factor α-induced protein 5; PTX3 is an acute phase protein that is generally expressed at a low level under stable conditions, but increases rapidly when inflammation is stimulated. Studies have shown that the plasma PTX3 level in patients with systemic lupus erythematosus is significantly higher than that in healthy people, and it is a marker for local inflammatory response to vascular damage in systemic lupus erythematosus. In some other rheumatic diseases, the PTX3 level is significantly higher than that in healthy people.

[0009] Soluble growth stimulatory gene 2 protein (sST2), serum soluble ST2 can participate in the myocardial fibrosis process after acute myocardial infarction, has diagnostic value for heart failure in patients with acute myocardial infarction, and can help judge the treatment effect. For patients with acute myocardial infarction after percutaneous coronary intervention, serum sST2 can improve the predictive value of adverse cardiovascular events.

[0010] Commonly used methods for detecting inflammatory factors include: enzyme-linked immunosorbent assay (ELISA), immunoblotting (WB), and flow cytometry (FACS). Among them, ELISA is the most common in clinical testing, with high sensitivity and good specificity, but it has many shortcomings in the clinical testing process, such as single detection target, excessive amount of detection samples, and low detection throughput; immunoblotting (WB) and its derived immunoblotting chip (WB array) have the disadvantages of large amount of detection samples and long detection time.

[0011] Cytometric Bead Array (CBA) liquid-based chip technology (hereinafter referred to as CBA) is a detection method based on the capture of labeled microspheres and the detection of patient body fluid targets by flow cytometry. It can simultaneously perform qualitative and quantitative detection of multiple indicators in the sample; in addition. It has the advantages of small sample volume, high sensitivity, good stability, short detection process, simple operation, and high detection rate. At present, its detection method has been widely used in the fields of cell biology, immunology, antibody drug screening, etc., providing data support for life science research. Therefore, the development of an inflammatory factor detection based on CBA technology is of great significance for practical applications. Summary of the invention

[0012] The technical problem to be solved by the present invention is that the detection time of the detection kit in the prior art is long and the detection steps are complicated.

[0013] In order to solve the above technical problems, the present invention provides an inflammatory factor detection kit based on liquid-based chip technology, wherein the inflammatory factor detection kit comprises: 6 fluorescent microspheres coupled to capture antibodies, and 6 detection antibodies labeled with reporter fluorescence; among the 6 fluorescent microspheres containing capture antibodies, the fluorescence intensity of each fluorescent microsphere is different.

[0014] The detection kit provided by the present invention is based on liquid-based chip technology, uses fluorescein to fluorescently encode microspheres, and connects capture antibodies to fluorescent microspheres through chemical reactions; the detection target is captured by the fluorescent microspheres, and the signal is displayed by the detection antibody, thereby completing the detection of inflammatory factors. This solves the previous problems of large sample demand for soluble inflammatory factor detection, long detection time and cumbersome detection steps, and greatly facilitates scientific researchers at the hospital and industrial ends.

[0015] In the present invention, each fluorescent microsphere is fluorescently encoded by fluorescent pigments of different concentrations, and one fluorescently encoded microsphere corresponds to one detection target.

[0016] Compared with the existing four-item detection method for inflammatory factors, the method provided in this application provides six tests.

[0017] Preferably, the fluorescent microspheres are microspheres treated with fluorescein encoding.

[0018] Preferably, the fluorescein is allophycocyanin dye (APC).

[0019] Preferably, the particle size of the fluorescent microspheres is 3 to 7 μm, for example, 3 μm, 4 μm, 5 μm, 6 μm or 7 μm.

[0020] Preferably, the six types of fluorescent microspheres coupled to capture antibodies are: fluorescent microspheres coupled to sCD25 monoclonal antibodies, fluorescent microspheres coupled to sCD40L monoclonal antibodies, fluorescent microspheres coupled to sCD130 monoclonal antibodies, fluorescent microspheres coupled to sTREM-1 monoclonal antibodies, fluorescent microspheres coupled to PTX3 monoclonal antibodies, and fluorescent microspheres coupled to sST2 monoclonal antibodies.

[0021] Preferably, the reporter fluorescence is phycoerythrin dye (PE).

[0022] Preferably, the six types of detection antibodies labeled with reporter fluorescence are: sCD25 monoclonal antibody labeled with reporter fluorescence, sCD40L monoclonal antibody labeled with reporter fluorescence, sCD130 monoclonal antibody labeled with reporter fluorescence, sTREM-1 monoclonal antibody labeled with reporter fluorescence, PTX3 monoclonal antibody labeled with reporter fluorescence and sST2 monoclonal antibody labeled with reporter fluorescence.

[0023] The PTX3 monoclonal antibody and sST2 monoclonal antibody used in this application are currently mainly used in enzyme-linked immunosorbent assays, and are not used based on CBA technology. The existing enzyme-linked immunosorbent assay has lower sensitivity than CBA technology. In addition, the target body expression levels recognized by the two antibodies, PTX3 monoclonal antibody and sST2 monoclonal antibody, are relatively low. It is precisely because of the application of CBA technology that the two targets can be better displayed at low concentrations.

[0024] In addition, the existing enzyme-linked immunosorbent assay can only detect one target at a time, while the present application is based on CBA technology and can detect more than four targets at a time.

[0025] In this application, there is only one capture antibody on the surface of the fluorescent microsphere group with the same concentration of fluorescent labeling. The capture antibody specifically binds to the target protein to be detected during the sample incubation stage, and then a detection antibody labeled with PE dye is added to continue incubation. After the detection antibody specifically binds to the target protein, the target protein that is not bound to the microsphere is washed away by washing buffer. Single proteins are identified by specific microsphere-fluorescence combinations, and the PE fluorescence level reflects the amount of protein captured on the microsphere. The PE signal generated by the standard is compared with the standard curve generated by the signal generated by the purified protein standard of known concentration to determine the content of the target protein to be measured.

[0026] Preferably, the inflammatory factor detection kit further comprises a freeze-dried detection target standard mixture, a sample diluent and a washing buffer.

[0027] Preferably, the detection instrument used in the inflammatory factor detection kit is a flow cytometer having a 488nm laser and a 638nm laser and 6 fluorescence channels.

[0028] Flow cytometry can perform high-speed quantitative measurements of multiple fluorescent microspheres simultaneously.

[0029] The implementation of the present invention has the following beneficial effects:

[0030] 1. The inflammatory factor detection kit of the liquid-based chip technology of the present application is a detection reagent using the CBA method, which distinguishes different targets by the difference in APC fluorescence intensity; each group of fluorescent microspheres has a unique capture antibody label, so that during the detection process, it can quickly, accurately and specifically bind to a specific inflammatory factor target. A single reaction system can detect multiple targets and can be repeated for multiple times, which greatly improves the detection throughput and reduces the difficulty of detecting inflammatory factors at the hospital or industrial end;

[0031] 2. The inflammatory factor detection kit of the liquid-based chip technology of the present application can be adapted to most two-light six-color flow cytometers on the market, and has high versatility;

[0032] 3. Compared with ELISA and immunoblotting, the single sample volume of the inflammatory factor detection kit described in this application is only 25 μL, while the single sample volume of ELISA and immunoblotting is 200 μL or even higher. This can greatly reduce the sample volume required for detection;

[0033] 4. Thanks to the CBA detection method, the well-to-well differences and CV values ​​of the kit are small;

[0034] 5. The inflammatory factor detection kit using liquid-based chip technology of the present application can avoid the fluorescence crosstalk caused by fluorescence overlap in current detection methods, thereby greatly improving the sensitivity and accuracy of detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is a schematic diagram of the detection of the kit provided by the present invention.

[0036] Figure 2 This is a light flow diagram of the fluorescent microspheres in the present invention in the front side.

[0037] Figure 3 This is the flow cytometric graph of fluorescent microspheres with different targets in the allophycocyanin channel. DETAILED DESCRIPTION

[0038] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0039] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0040] The sources of the capture antibodies in the following embodiments of the present invention are shown in Table 1 below:

[0041] Table 1

[0042] name source sCD25 Biolegend sCD40L Biolegend sCD130 Biolegend sTREM Biolegend PTX3 Biolegend sST2 Biolegend

[0043] The sources of the detection antibodies in the following embodiments of the present invention are shown in Table 2 below:

[0044] Table 2

[0045] name source CD25-PE Biolegend CD40L-PE Biolegend CD130-PE Biolegend TREM-PE Biolegend PTX3-PE Biolegend ST2-PE Biolegend

[0046] The sources of the protein standards in the following embodiments of the present invention are shown in Table 3 below:

[0047] Table 3

[0048]

[0049]

[0050] In the following examples of the present application: 5 μm microspheres are from the microspheres produced by NanoMicro, and the capture antibodies are from the antibodies in Table 1 above. The freeze-dried protein standard is from Biolegend, and the detection antibodies are all the detection target antibodies labeled with PE dye.

[0051] The present application discloses a kit for detecting inflammatory factors based on liquid-based chip technology. The specific method for performing the detection is as follows. See the schematic diagram. Figure 1 As shown:

[0052] 1. Prepare samples: Serum or plasma samples do not need to be diluted. Cell culture medium and cell culture supernatant need to be diluted with sample diluent. The recommended dilution multiple is 2-4 times.

[0053] 2. Preparation before the experiment: Centrifuge the fluorescent microspheres with fixed capture antibodies under light-proof conditions; the centrifugation time is 1 min;

[0054] 3. Preparation of standard: After centrifugation at 12000rpm for 1min, the freeze-dried standard mixture was dissolved with sample diluent and marked as S1. Then, 7 1.5mL ep tubes were taken and marked as S2-S8 respectively. 25μL of S1 was transferred to S2 using a pipette and shaken evenly. Then, 25μL of S2 was transferred to S3 and shaken evenly. The rest of S4-S7 were processed according to the above method. Sample diluent was added to S8 tube as a control.

[0055] 4. Experimental process: Add the fluorescent microsphere mixture connected with monoclonal antibodies of sCD25, sCD40L, sCD130, sTREM-1, PTX3, and sST2 into the flow tube according to the experimental requirements. Then, add the standard dilution gradient and the test sample into the flow tube, and then add the detection antibody in turn, and incubate at room temperature for 2 hours; after centrifugation to remove the supernatant, add washing buffer to wash the fluorescent microspheres. After washing, add 200 μL washing buffer, and then analyze on the flow cytometer.

[0056] 5. Data acquisition and analysis: Flow cytometry was used to acquire data, and the data were analyzed using analysis software (Flowjo, FCAP).

[0057] Example 1

[0058] The inflammatory factor detection kit provided by the present application with liquid-based chip technology includes the following components:

[0059] A mixture of 5 μm particle size, fluorescent microspheres encoded by fluorescein coupled to capture antibody sCD25, fluorescent microspheres encoded by fluorescein coupled to capture antibody sCD40L, fluorescent microspheres encoded by fluorescein coupled to capture antibody sCD130, fluorescent microspheres encoded by fluorescein coupled to capture antibody sTREM-1, fluorescent microspheres encoded by fluorescein coupled to capture antibody PTX3, and fluorescent microspheres encoded by fluorescein coupled to capture antibody sST; sample diluent (PBS solution of 0.05% Tween-20, 0.1% BSA (bovine serum albumin)), washing buffer (PBS solution of 0.05% Tween-20);

[0060] The detection antibodies consisted of the following: PE-labeled sCD25 monoclonal antibody, PE-labeled sCD40L monoclonal antibody, PE-labeled sCD130 monoclonal antibody, PE-labeled sTREM-1 monoclonal antibody, PE-labeled PTX3 monoclonal antibody, and PE-labeled sST2 monoclonal antibody;

[0061] Lyophilized protein standards (as shown in Table 3).

[0062] The kit provided in the above examples was subjected to performance evaluation. The performance evaluation included sensitivity, standard sample recovery rate, and intra-batch and inter-batch precision. The experimental steps were as follows:

[0063] 1. Sample preparation: Serum or plasma samples do not need to be diluted. Cell culture medium and cell culture supernatant need to be diluted with sample diluent. The recommended dilution multiple is 2 times.

[0064] 2. Preparation before the experiment: Centrifuge the fluorescent microspheres with fixed capture antibodies under light-proof conditions; the centrifugation time is 1 min;

[0065] 3. Prepare the standard: After centrifuging the freeze-dried standard mixture at 12000rpm for 1min, dissolve it with sample diluent and mark it as S1. Then take 7 1.5mL ep tubes and mark them as S2-S8 respectively. Use a pipette to transfer 25μL of S1 to S2 and shake evenly. Then take 25μL of S2 to S3 and shake evenly. The rest of S4-S7 are processed according to the above method. Add sample diluent to S8 tube as a control;

[0066] 4. Experimental process: Add the fluorescent microspheres connected with the capture antibody into the flow tube according to the experimental requirements, then add the standard dilution gradient and the test sample into the flow tube, then add the detection antibody in sequence, and incubate at room temperature for 2 hours; after centrifugation to remove the supernatant, add the washing buffer to wash the fluorescent microspheres, add 200μL washing buffer after washing, and then analyze on the flow cytometer;

[0067] 5. Data acquisition and analysis: Set up APC and PE detection channels in the flow cytometer, and set FSC (forward scatter) in the flow cytometer software interface ( Figure 2 ), SSC (side scatter) graph, APC channel and PE channel cross analysis graph ( Figure 3 ). First, the S8 sample was put on the machine, the voltage was adjusted in the FSC / SSC graph, and the initial microsphere group was circled and gated. Then, the corresponding targets were circled and gated according to the microsphere fluorescence coding and the microsphere fluorescence intensity in the flow graph. The PE channel voltage was adjusted so that the MFI (mean fluorescence intensity) of the PE channel of all detection targets was around 0. Then all samples were tested on the machine, and the total number of microspheres read was 1800. All data were exported as FCS files and analyzed in the corresponding software.

[0068] In the above tests, the corresponding relationship between the gate and the corresponding target protein is shown in Table 4:

[0069] Table 4

[0070] Gating Target protein P1 CD25 P2 CD40L P3 CD130 P4 sTREM P5 PTX3 P6 ST2

[0071] 1. Sensitivity test

[0072] According to the above experimental process, each target protein standard was diluted with sample diluent, and the sensitivity test (detection limit) of the inflammatory factor detection kit was performed. The detection limit test results are shown in Table 5 below:

[0073] Table 5

[0074]

[0075]

[0076] 2. Cross-reactivity

[0077] No detectable or significant cross-reactivity was detected between targets.

[0078] 3. Calibration recovery rate

[0079] The target protein was diluted to a fixed concentration, and the standard diluted to a fixed concentration was tested according to the above experimental steps, and the test concentration was compared with the theoretical concentration to calculate the recovery rate. The analysis results are shown in Table 6 below:

[0080] Table 6

[0081] Target protein Recovery rate (%) CD25 95.3 CD40L 102.5 CD130 99.3 sTREM 98.7 PTX3 110.5 ST2 102.8

[0082] 4. Intra-batch precision

[0083] The target protein was diluted to a medium concentration and a lower concentration using a sample diluent, and the test was performed according to the above experimental method. A single concentration was repeated 10 times to obtain the intra-batch precision or coefficient of variation (CV) of the sample. The data results are shown in Table 7.

[0084] Table 7

[0085]

[0086]

[0087] 5.Batch-to-batch precision

[0088] The target protein was diluted to a medium concentration and a lower concentration using a sample diluent, and the test was performed according to the above experimental method. A single concentration was repeated 10 times to obtain the intra-batch precision or coefficient of variation (CV) of the sample. The data results are shown in Table 8.

[0089] Table 8

[0090]

[0091] It can be seen from the above test results that the kit provided in this application has a low detection limit concentration, among which the target protein concentration with the highest detection limit is only 3.02pg / mL (TREM), the detection sensitivity is high, and there is no cross-reactivity between targets.

[0092] Through the calculation and analysis of intra-batch precision and inter-batch precision, the detection performance of the kit provided in this application is stable, and it is a stable and reliable detection method.

[0093] Compared with existing detection methods, the present invention has high detection sensitivity, short detection time and detection steps, and is more conducive to practical application.

[0094] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An inflammatory factor detection kit based on liquid-based chip technology, characterized in that: The inflammatory factor detection kit comprises: 6 fluorescent microspheres coupled with capture antibodies and 6 detection antibodies labeled with reporter fluorescence; among the 6 fluorescent microspheres containing capture antibodies, the fluorescence intensity of each fluorescent microsphere is different.

2. The inflammatory factor detection kit according to claim 1, characterized in that: The fluorescent microspheres are microspheres treated with fluorescein encoding.

3. The inflammatory factor detection kit according to claim 2, characterized in that: The fluorescein is an allophycocyanin dye.

4. The inflammatory factor detection kit according to claim 2, characterized in that: The particle size of the fluorescent microspheres is 3 to 7 μm.

5. The inflammatory factor detection kit according to claim 1, characterized in that: The six types of fluorescent microspheres coupled with capture antibodies are: fluorescent microspheres coupled with sCD25 monoclonal antibody, fluorescent microspheres coupled with sCD40L monoclonal antibody, fluorescent microspheres coupled with sCD130 monoclonal antibody, fluorescent microspheres coupled with sTREM-1 monoclonal antibody, fluorescent microspheres coupled with PTX3 monoclonal antibody and fluorescent microspheres coupled with sST2 monoclonal antibody.

6. The inflammatory factor detection kit according to claim 1, characterized in that: The reporter fluorescence is phycoerythrin dye.

7. The inflammatory factor detection kit according to claim 1, characterized in that: The six reporter fluorescence-labeled detection antibodies are: reporter fluorescence-labeled sCD25 monoclonal antibody, reporter fluorescence-labeled sCD40L monoclonal antibody, reporter fluorescence-labeled sCD 130 monoclonal antibody, reporter fluorescence-labeled sTREM-1 monoclonal antibody, reporter fluorescence-labeled PTX3 monoclonal antibody and reporter fluorescence-labeled sST2 monoclonal antibody.

8. The inflammatory factor detection kit according to claim 1, characterized in that: The inflammatory factor detection kit also includes a freeze-dried detection target standard mixture, a sample diluent and a washing buffer.

9. The inflammatory factor detection kit according to claim 1, characterized in that: The detection instrument used in the inflammatory factor detection kit is a flow cytometer with a 488nm laser and a 638nm laser and 6 fluorescence channels.

Images