N-carbohydrate chain marker for predicting seizure part of type 2 hereditary angioedema, kit and application of N-carbohydrate chain marker

Abstract

The invention relates to the technical field of medical detection, in particular to an N-carbohydrate chain marker for predicting a seizure site of type 2 hereditary angioedema, a kit and application of the N-carbohydrate chain marker. In the N-carbohydrate chain marker provided by the invention, the contents of H6N5F1E2L1 and H5N4F1E1L1 are remarkably changed in the body of a patient suffering from laryngeal edema, and H3N3F1, H6N3E1 and H5N2 are remarkably changed in the body of a patient suffering from abdominal edema, so that the N-carbohydrate chain marker shows relatively good seizure site identification capability of type 2 hereditary angioedema, and the seizure site identification capability of type 2 hereditary angioedema is remarkably improved. The kit has high specificity, sensitivity and accuracy. The N-carbohydrate chain marker has the advantages of being convenient to detect, short in required time and the like when being used for predicting the site of attack, and has important significance for giving personalized prevention and treatment measures in time.

Description

Technical Field

[0001] The present invention relates to the field of medical detection technology, and in particular to an N-sugar chain marker for predicting the onset site of hereditary angioedema type 2, a kit and applications thereof. Background Art

[0002] Hereditary angioedema (hAE) is a rare, life-threatening genetic disease with an incidence of approximately 1 / 50,000. The clinical manifestations are highly heterogeneous and are characterized by acute, recurrent subcutaneous and / or submucosal tissue edema, which can affect the face, limbs, upper respiratory tract, and gastrointestinal tract. Classic HAE includes HAE type 1 (HAE-1) and HAE type 2 (HAE-2), among which the incidence of HAE-2 is much lower than that of HAE-1. At present, the diagnosis of HAE internationally requires a combination of family history, clinical manifestations, and laboratory tests. However, due to the rarity of HAE, clinical heterogeneity, and defects in existing diagnostic markers, patients are often misdiagnosed and mistreated, resulting in delayed diagnosis of HAE.

[0003] Due to the rarity of HAE, doctors and the public are not well aware of HAE, resulting in the vast majority of HAE patients going to multiple hospitals for many years but still not getting the correct diagnosis and treatment. The clinical heterogeneity of HAE is specifically reflected in the age of onset, frequency of attacks, triggering factors of acute attacks, prodromal signs, severity of edema and predilection sites. Moreover, the symptoms of each attack of the same patient are also different. In mild cases, only mild acral edema may be manifested, which does not affect normal daily life. In severe cases, if the gastrointestinal tract is involved, severe abdominal pain, nausea, vomiting, diarrhea, and abdominal effusion may occur. It is often misdiagnosed as acute abdomen, leading to unnecessary abdominal surgery. If respiratory mucosal edema occurs, it can quickly cause dyspnea or suffocation, causing some patients to undergo multiple tracheotomies due to repeated attacks of laryngeal edema. Some patients suffocate to death due to untimely rescue. More than 50% of HAE patients will experience laryngeal edema and are at risk of death. HAE patients are often in a state of severe anxiety and fear, which leads to insufficient or excessive prevention, greatly affecting the patients' quality of life and life expectancy, and increasing the economic burden.

[0004] Due to the unpredictability of HAE edema attacks and the lack of predictive markers for the time and location of edema attacks, even if a clear diagnosis of HAE is made, it is still impossible to predict the time, location and severity of the patient's edema attacks, so it is impossible to provide personalized prevention and treatment measures. Therefore, there is an urgent need for specific biomarkers to guide doctors to identify the risk of laryngeal edema earlier, so that timely treatment measures can be taken, such as specific drugs or tracheotomy, to save the patient's life. At the same time, through marker prediction and personalized prevention and treatment, it helps to reduce the frequency and severity of laryngeal edema, thereby improving the quality of life of patients. Chinese patent application CN113311056A discloses markers for the diagnosis of laryngeal edema in hereditary angioedema, but it is for HAE-1 type rather than HAE-2 type.

[0005] Protein glycosylation is one of the most common and complex post-translational modifications. The glycans on proteins mainly play a role by participating in a variety of important biological processes such as cell adhesion, protein folding, molecular transport and clearance, receptor activation and signal transduction. Therefore, protein glycosylation can significantly affect the structure and function of proteins. Protein glycosylation can be divided into N-glycosylation, O-glycosylation, C-mannosylation and glycosylphosphatidylinositol anchoring, of which more than half of glycoproteins are N-glycosylated. In recent years, the development of glycomics technology has enabled a more comprehensive understanding of the biological significance of protein glycosylation in physiological and disease states. Glycomics has a more prominent diagnostic and predictive ability in many aspects such as disease diagnosis, prediction of progression and prognosis. Studies have found that changes in the N-glycan spectrum in plasma are related to the occurrence and development of many diseases, such as lung cancer, colorectal cancer and other cancers, Alzheimer's disease, type 2 diabetes cardiovascular disease, etc. At present, the N-glycan markers related to the site of hereditary angioedema are still relatively limited, and there is an urgent need to develop N-glycan markers with strong predictive ability. Summary of the invention

[0006] The present invention provides an N-sugar chain marker for predicting the onset site of type 2 hereditary angioedema, a kit and applications thereof.

[0007] Specifically, the present invention provides the technical solution described below.

[0008] In a first aspect, the present invention provides markers for predicting the onset site of hereditary angioedema type 2, wherein the markers include a combination of one or more of the following sugar chain markers: H6N5F1E2L1, H5N4F1E1L1, H3N3F1, H6N3E1 and H5N2.

[0009] The present invention develops sugar chain markers with predictive significance for the onset site of hereditary angioedema type 2, screens N-sugar chain markers related to the onset of laryngeal edema and abdominal edema by detecting and analyzing the N-sugar chain composition and spectral structure of patients with hereditary angioedema type 2 who have onset of laryngeal edema and abdominal edema, and determines markers with potential for clinical predictive application through methods such as ROC curve analysis.

[0010] Among the above markers, the expression levels of H6N5F1E2L1 and H5N4F1E1L1 were significantly changed in patients with type 2 hereditary angioedema who had laryngeal edema attacks. The AUC values ​​for distinguishing between patients without laryngeal edema and patients with laryngeal edema reached above 0.97. They can accurately distinguish between patients without laryngeal edema and patients with laryngeal edema, and have high specificity and sensitivity for predicting whether patients with type 2 hereditary angioedema will have laryngeal edema attacks.

[0011] Among the above markers, the expression levels of H3N3F1, H6N3E1 and H5N2 were significantly changed in patients with type 2 hereditary angioedema who had abdominal edema attacks. The AUC values ​​for distinguishing between patients without abdominal edema and patients with abdominal edema attacks were all above 0.8. They can accurately distinguish between patients without abdominal edema and patients with abdominal edema attacks, and have high specificity and sensitivity for predicting whether patients with type 2 hereditary angioedema will have abdominal edema attacks.

[0012] The above five sugar chain markers can be used alone to predict the site of onset of hereditary angioedema type 2. The onset site is specifically the throat or abdomen. Among them, H6N5F1E2L1 and H5N4F1E1L1 are used to predict whether the onset site of edema is the throat (i.e., whether laryngeal edema will occur), and H3N3F1, H6N3E1 and H5N2 are used to predict whether the onset site of edema is the abdomen (i.e., whether abdominal edema will occur).

[0013] In some embodiments of the present invention, the marker comprises any one of the following sugar chain markers: H6N5F1E2L1, H5N4F1E1L1, H3N3F1, H6N3E1, H5N2.

[0014] Since the AUC value of any one of the above five sugar chain markers in predicting the occurrence site of hereditary angioedema type 2 is above 0.8, the AUC value of any 2, 3, 4, or 5 of them used in combination for predicting the occurrence site of hereditary angioedema type 2 must also be above 0.8, or even higher than the AUC value of each individual marker in the combination. Therefore, a combination of at least two of the above sugar chain markers can also be used to predict the occurrence site of hereditary angioedema type 2 with high accuracy.

[0015] In some embodiments of the present invention, the marker comprises a combination of at least two of the following sugar chain markers: H6N5F1E2L1, H5N4F1E1L1, H3N3F1, H6N3E1, H5N2.

[0016] In some specific embodiments of the present invention, a marker for predicting the onset site of hereditary angioedema type 2 is provided, wherein the marker comprises one or a combination of two of the following sugar chain markers: H6N5F1E2L1, H5N4F1E1L1. The onset site is the larynx. The above marker can be used to predict whether a patient with hereditary angioedema type 2 will develop laryngeal edema.

[0017] In some specific embodiments of the present invention, a marker for predicting the onset site of hereditary angioedema type 2 is provided, wherein the marker comprises a combination of one or more of the following sugar chain markers: H3N3F1, H6N3E1, and H5N2. The onset site is the abdomen. The above marker can be used to predict whether a patient with hereditary angioedema type 2 will develop abdominal edema.

[0018] Preferably, in patients with type 2 hereditary angioedema who have developed laryngeal edema, the expression levels of H6N5F1E2L1 and H5N4F1E1L1 are significantly increased compared to those in patients with type 2 hereditary angioedema who have never developed laryngeal edema.

[0019] Preferably, in patients with type 2 hereditary angioedema who have abdominal edema, the expression level of H3N3F1 is significantly increased compared with that in patients with type 2 hereditary angioedema who have never had abdominal edema; the expression levels of H6N3E1 and H5N2 are significantly decreased compared with that in patients with type 2 hereditary angioedema who have never had abdominal edema.

[0020] The naming method of the sugar chain markers described in the present invention refers to the following literature: Zhang Z, Westhrin M, Bondt A, et al. Serum protein N-glycosylation changes in multiple myeloma[J]. Biochimica et Biophysica Acta (BBA)-General Subjects, 2019 May; 1863(5): 960-970. The sugar chain markers of the present invention are all sugar chain structures directly detected by mass spectrometry.

[0021] In the present invention, the nomenclature and molecular weight of the directly detected sugar chain structure are shown in Table 1.

[0022] Table 1

[0023] In a second aspect, the present invention provides the use of the above-mentioned marker for predicting the onset site of hereditary angioedema type 2 or its detection reagent in the preparation of a product for predicting the onset site of hereditary angioedema type 2.

[0024] Preferably, the product is a medicine or a kit.

[0025] In some embodiments of the present invention, provided is a marker for predicting the onset site of hereditary angioedema type 2 or a detection reagent thereof in the preparation of a product for predicting whether the onset site of hereditary angioedema type 2 is the larynx, wherein the marker comprises H6N5F1E2L1 and / or H5N4F1E1L1.

[0026] Among them, it is predicted whether the onset site of hereditary angioedema type 2 is the larynx, that is, whether laryngeal edema will occur.

[0027] In some embodiments of the present invention, a marker for predicting the onset site of hereditary angioedema type 2 or a detection reagent thereof is provided for use in preparing a product for predicting whether the onset site of hereditary angioedema type 2 is the abdomen, wherein the marker comprises one or more selected from H3N3F1, H6N3E1, and H5N2.

[0028] Among them, it is predicted whether the onset site of hereditary angioedema type 2 is the abdomen, that is, whether abdominal edema will occur.

[0029] In a third aspect, the present invention provides use of the above-mentioned markers for predicting the onset site of hereditary angioedema type 2 as drug targets for preventing or treating hereditary angioedema type 2.

[0030] In a fourth aspect, the present invention provides a product for predicting the onset site of hereditary angioedema type 2, wherein the product comprises the above-mentioned marker for predicting the onset site of hereditary angioedema type 2 or a detection reagent thereof.

[0031] Preferably, the product is a medicine or a kit.

[0032] In the present invention, the detection reagent of the marker can be any reagent used to detect the marker of the present invention, for example: characteristic sugar chain probe, mass spectrometry detection reagent, lectin chip, etc.

[0033] In some embodiments of the present invention, the detection reagent is a characteristic sugar chain probe that specifically recognizes the sugar chain marker.

[0034] In some embodiments of the present invention, the detection reagent is a mass spectrometry detection reagent for detecting the sugar chain marker.

[0035] In a fifth aspect, the present invention provides a device for predicting the onset site of hereditary angioedema type 2, the device comprising: A detection module, used to detect the content of the above-mentioned marker for predicting the onset site of hereditary angioedema type 2 in the sample to be tested; An input module is used to obtain the detection result of the detection module; A judgment module, used for predicting the onset site of hereditary angioedema type 2 according to the test results obtained by the input module; Output module, used to output prediction results.

[0036] Preferably, the judgment standard of the judgment module is: if the content of H6N5F1E2L1 and / or H5N4F1E1L1 in the sample to be tested is significantly higher than that in type 2 hereditary angioedema patients who have never had laryngeal edema, it is predicted that the site of edema onset is the larynx; if the content of H3N3F1 in the sample to be tested is significantly higher than that in type 2 hereditary angioedema patients who have never had abdominal edema, and / or the expression levels of H6N3E1 and H5N2 are significantly lower than those in type 2 hereditary angioedema patients who have never had abdominal edema, it is predicted that the site of edema onset is the abdomen.

[0037] Preferably, the sample to be tested is plasma.

[0038] In a sixth aspect, the present invention provides a method for predicting the onset site of hereditary angioedema type 2, the method comprising: detecting the content of the above-mentioned marker for predicting the onset site of hereditary angioedema type 2 in the subject's plasma, and predicting the onset site of hereditary angioedema type 2 based on the content of the marker.

[0039] Preferably, if the content of H6N5F1E2L1 and / or H5N4F1E1L1 is significantly higher than that of type 2 hereditary angioedema patients who have never experienced laryngeal edema, it is predicted that the site of edema onset is the larynx, i.e., laryngeal edema occurs; if the content of H3N3F1 in the sample to be tested is significantly higher than that of type 2 hereditary angioedema patients who have never experienced abdominal edema, and / or the expression levels of H6N3E1 and H5N2 are significantly lower than those of type 2 hereditary angioedema patients who have never experienced abdominal edema, it is predicted that the site of edema onset is the abdomen, i.e., abdominal edema occurs.

[0040] The beneficial effects of the present invention include at least: among the N-sugar chain markers provided by the present invention, the contents of H6N5F1E2L1 and H5N4F1E1L1 are significantly changed in patients with laryngeal edema, and H3N3F1, H6N3E1 and H5N2 are significantly changed in patients with abdominal edema. According to the ROC curve verification, the above-mentioned N-sugar chain markers all show very good ability to identify the attack site of type 2 hereditary angioedema, and can be used as a marker for predicting the attack site of type 2 hereditary angioedema, with high specificity, sensitivity and accuracy. The use of the N-sugar chain markers of the present invention for predicting the attack site also has the advantages of convenient detection and short time required, and can be used in practice to predict the attack site of type 2 hereditary angioedema, which is of great significance for timely personalized prevention and treatment measures. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0042] Figure 1 3 and 4. The ROC curves of the N-glycan markers in Example 1 of the present invention in predicting the occurrence of laryngeal edema (a, b) and abdominal edema (c, d) in hereditary angioedema type 2. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0044] The "glycome" mentioned in the present invention refers to all sugar chains expressed in a sample (such as body fluids, cells, tissues) or all sugar chains on a certain type of glycoprotein.

[0045] The sample of the present invention is selected from: body fluid samples, such as blood, serum, plasma, urine, saliva, cerebrospinal fluid, lymph, spinal fluid, ascites, amniotic fluid; cell samples, such as cell samples separated from tissues, cell samples cultured in vitro; tissue samples, which can be in the form of fresh tissue samples, fixed tissue samples, etc. In specific detection practice, plasma samples are preferred.

[0046] The sugar chains described in the present invention may be free sugar chains or sugar chains released from sugar complexes.

[0047] Free sugar chains can be obtained by techniques known in the art, including but not limited to: enzymatic methods, such as using glycosidase, preferably glycosidase PNGase F; chemical methods, such as using β-elimination reaction, glycoprotein hydrazinolysis reagent; and a combination of enzymatic and chemical methods can also be used to release sugar chains.

[0048] The derivatization methods of the present invention include but are not limited to: methyl amination, esterification, methylation, reductive amination, acetylation, etc. The type of derivatization can be selected according to the needs, and esterification is preferred.

[0049] In the present invention, after the sugar chains are released from the body fluid protein, the N-sugar chains can be purified and / or enriched using techniques known in the art. The purification and enrichment methods include, but are not limited to, centrifugation, filtration, extraction, adsorption, capillary electrophoresis, chromatography, and the like.

[0050] In a specific embodiment of the present invention, a CottonhILIC SPE separation column is used to enrich and purify N-glycans, wherein water is used to activate the separation column, a solution of water:acetonitrile = 15:85 (volume ratio) is used to equilibrate the separation column, and pure water is used to elute the sugar chains.

[0051] In the present invention, sugar chain analysis and data processing can be performed by using analytical methods known in the art to measure and quantify sugar groups. The methods include, but are not limited to, mass spectrometry, such as matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) (such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS), matrix-assisted laser desorption ionization-quadrupole ion trap-time of flight mass spectrometry (MALDI-QIT-TOF MS)), fast atom bombardment mass spectrometry (FAB-MS), electrospray mass spectrometry (ESI-MS), multistage mass spectrometry, high performance liquid chromatography HPLC, liquid chromatography-mass spectrometry (LC-MS), sugar chip technology, nuclear magnetic resonance NMR, or any combination of the above methods. Preferably, a high-resolution technique is used for analysis, such as MALDI MS.

[0052] In the present invention, the sugar chain analysis data is further calculated and processed to obtain the required sugar group related information. For example, the ratio of the peak area of ​​each sugar chain in the sample to the sum of all peak areas can be obtained, thereby obtaining the relative quantitative value of each sugar chain, which can avoid the deviation caused by the pretreatment of parallel samples and other operation processes, and ensure the high accuracy of the analysis; these data can be directly used for relative content comparison or qualitative analysis, and used to monitor the abundance changes of target sugar chains.

[0053] Sugar chain analysis data can be further calculated and processed using various related sugar chain analysis software, databases, algorithms, etc. The available sugar chain analysis software includes but is not limited to: MassyTools, Progenesis MALDI, LassyTools, GlycoWorkbench, GlycanMass, GlycoMod, GlycoFragment, GlycoSearchMS, etc. Available sugar chain analysis databases include but are not limited to: CCSD, GlycomeDB, CarbBank, EUROCarbDB, etc.

[0054] In the present invention, the sugar chain detection method is preferably a high-throughput detection method, for example, 96, 192, 288, 384 or more samples can be processed and detected simultaneously, which greatly reduces the time for sample preparation.

[0055] In the following examples, all HAE-2 patients were tested for blood sugar group. The cohort characteristics of the samples used are shown in Table 2. Among them, the site of edema attack in HAE-2 patients is shown in Table 3. It should be noted that HAE is a rare disease. Among HAE patients in China, HAE-1 accounts for 98.73%, while HAE-2 accounts for only 1.27% (Zhi Yuxiang, An Lixin, Lai He, et al. Expert consensus on the diagnosis and treatment of hereditary angioedema [J]. Chinese Journal of Clinical Immunology and Allergy, 2019, 13(1):4). Therefore, the number of clinical HAE-2 cases is relatively limited. For extremely rare diseases, the above sample population is representative, and the subsequent ROC curve analysis determined that the markers of the present invention have good predictive ability for the site of attack of type 2 hereditary angioedema.

[0056] Table 2 Baseline demographic information of hAE-2 patients

[0057] Table 3 Edema attack sites in hAE-2 patients

[0058] The above samples were used as research objects to screen plasma N-glycan markers for predicting the onset of edema in HAE-2 patients and determine their clinical application value. The specific steps are as follows: 1. Glycosidase releases N-glycans N-glycans were released from whole serum / plasma glycoproteins using glycosidase PNGase F. The specific steps were as follows: 5 μL serum / plasma was taken from each sample, 10 μL 2% SDS was added, and the mixture was incubated at 60°C for 10 minutes; then 10 μL enzymatic solution (containing 2% NP-40, 2.5 × PBS and 1 U PNGase F) was added and the mixture was incubated at 37°C for 12-16 hours.

[0059] 2. N-Glycan Derivatization The free N-glycans were derivatized using known derivatization techniques, and the sialic acids of α2,3 and α2,6 linkages could be distinguished after derivatization. The specific steps are as follows: 1 μL of the above enzymatically hydrolyzed serum / plasma, add 20 μL of derivatization reagent (250 mM EDC and 250 mMhOBt, the solvent is anhydrous ethanol), and incubate at 37°C for 60 minutes.

[0060] 3. N-glycan HILIC-SPE enrichment and purification The derivatized sugar chains obtained above were enriched and purified by HILIC-SPE. HILIC uses cotton thread as the stationary phase, and the cotton thread is filled in a 20μL pipette tip to make a purification column. First, the column is activated 3 times with 15μL ultrapure water (MQ); then, the column is equilibrated with 15μL of 85% acetonitrile (ACN) 3 times; then, the derivatized sugar chain mixture is added to the column and loaded 30 times to ensure that the derivatized N-sugar chains are adsorbed on the column as completely as possible; then, the column is eluted 3 times with 15μL of 85% acetonitrile + 1% trifluoroacetic acid (TFA), and then 15μL of 85% acetonitrile 3 times; finally, the sugar chains are eluted in 10μL MQ.

[0061] 4. Mass spectrometry analysis of N-glycans Before detection, the mass spectrometer was calibrated with a peptide mixture standard of known molecular mass (Bruker Peptide Calibration Standard II). The matrix super-DHB was dissolved in a 50% acetonitrile (water) solution containing 1 mM NaOH at a concentration of 5 mg / mL. Take 1 μL of the above purified N-glycans and spot them on the mass spectrometry target plate, then add 1 μL of the matrix solution on the sample and dry it at room temperature. MALDI-TOF MS was used for analysis. The mass spectrometer was equipped with a Smartbeam 3D laser source and collected signal ions in the positive ion reflection mode (reflection positive, RP). FlexControl software was used for control. The m / z range was set to 1000 to 5000 during sample detection. The spectrum acquisition setting was: for each sample point on the mass spectrometry target plate, the laser completely randomly collected signals within the sample point range, accumulated 10K laser shots, collected a mass spectrum, and the laser frequency was 5000 Hz.

[0062] 5. Data preprocessing and statistical analysis The collected mass spectra were preprocessed using FlexAnalysis and MassyTools software and exported to Microsoft Excel for further analysis. The sugar chain analysis function of GlycoWorkbench was used to assist manual analysis to analyze the mass spectrometry data. The identification of sugar chain structure was mainly based on mass-to-charge ratio, secondary mass spectrometry fragment attribution and published literature. The quantification of a single sugar chain was obtained by the peak area of ​​a single sugar chain / the sum of the peak areas of all sugar chains detected. All ions were in the form of sodium addition, i.e. [M+Na] + .H = hexose (mannose or galactose); N = N-acetylglucosamine (N-acetylglucosamine: GlcNAc); F = deoxyribose (fucose); L = α2,3-linked sialic acid; E = α2,6-linked sialic acid. The differences in N-glycosylation between HAE-2 patients with and without laryngeal and / or abdominal edema and the relationship between N-glycosylation characteristics and clinical parameters were evaluated by statistical tests, regression analysis, and receiver operating characteristic curves. The quality of mass spectrometry data for the study cohort was evaluated by the standards randomly distributed on the target plate during the sample testing process and the calculated mean, coefficient of variation, and standard deviation of each glycan for multiple standards.

[0063] 6. Results and Discussion The average CV value of the top 30 sugar chains obtained from the quality control standard was 9.749%, indicating that the data obtained in the present invention is reliable.

[0064] Example 1: Use of N-glycan markers to predict the onset of edema in HAE-2 patients A total of 109 glycan structures were detected in the HAE-2 edema attack site study cohort (Table 2), of which 63 glycans passed quality testing for subsequent analysis, with an average CV value of 5.17% for the top 10 glycans. The differences in the 63 directly detected N-glycans in HAE-2 patients with / without laryngeal edema or abdominal edema were first analyzed.

[0065] Among the 63 directly detected sugar chain features found above, 2 sugar chain features showed significant differences between HAE-2 patients with and without laryngeal edema. The results showed that compared with HAE-2 patients who had never experienced laryngeal edema, the levels of H6N5F1E2L1 and H5N4F1E1L1 were significantly increased in HAE-2 patients with laryngeal edema. According to the receiver operating characteristic (ROC) test results, it was found that the above 2 sugar chain features can effectively distinguish patients with or without laryngeal edema. Among the 63 directly detected sugar chain features, 3 sugar chain features showed significant differences between HAE-2 patients with and without abdominal edema. The results showed that compared with HAE-2 patients who had never experienced abdominal edema, the content of H3N3F1 was significantly increased in HAE-2 patients with abdominal edema, and the content of H6N3E1 and H5N2 was significantly decreased in HAE-2 patients with abdominal edema. According to the results of the receiver operating characteristic (ROC) test, it was found that the above three sugar chain features can effectively distinguish patients with or without abdominal edema attacks. These N-sugar chain features can be used as potential markers to predict the site of edema attacks in HAE-2 patients. Table 4 lists the relevant sugar chains of two groups of HAE-2 patients with laryngeal edema and those without laryngeal edema attacks, as well as the area under the curve (AUC) obtained by evaluating the above sugar chains for predicting the site of edema attacks in HAE-2 patients using the receiver operating characteristic (ROC) curve. Table 5 lists the relevant sugar chains of two groups of HAE-2 patients with and without abdominal edema attacks, as well as the area under the curve (AUC) obtained by evaluating the above sugar chains for predicting the site of edema attacks in HAE-2 patients using the receiver operating characteristic (ROC) curve. Among them, the ROC curves of H6N5F1E2L1 and H5N4F1E1L1 for predicting laryngeal edema in HAE-2 patients and the ROC curves of H3N3F1, H6N3E1, and H5N2 for predicting abdominal edema in HAE-2 patients are shown in Figure 5. Figure 1 shown.

[0066] Table 4

[0067] Note: AUC p value: p<0.05: diagnostic efficacy is significantly better than random p≥0.05: not statistically significant.

[0068] Table 5

[0069] Note: AUC p value: p<0.05: diagnostic efficacy is significantly better than random p≥0.05: not statistically significant.

[0070] In summary, the present invention found that in HAE-2 patients with laryngeal edema, H6N5F1E2L1 and H5N4F1E1L1 were significantly correlated with laryngeal edema. Further establishment of the ROC curve found that these two sugar chains showed very good discrimination ability, with AUCs of 1.000 and 0.971, respectively. In HAE-2 patients with abdominal edema, H3N3F1, H6N3E1, and H5N2 were significantly correlated with abdominal edema. Further establishment of the ROC curve found that these three sugar chains showed very good discrimination ability, with AUCs of 0.886, 0.857, and 0.829, respectively. The above markers have high specificity and sensitivity for predicting the site of edema onset in HAE-2 patients, and can be used alone or in combination to predict the site of edema onset in HAE-2 patients.

[0071] 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. A marker for predicting the onset site of hereditary angioedema type 2, characterized in that: The markers include a combination of one or more of the following sugar chain markers: H6N5F1E2L1, H5N4F1E1L1, H3N3F1, H6N3E1 and H5N2.

2. The marker for predicting the onset site of hereditary angioedema type 2 according to claim 1, characterized in that: In patients with type 2 hereditary angioedema who had laryngeal edema, the expression levels of H6N5F1E2L1 and H5N4F1E1L1 were significantly higher than those in patients with type 2 hereditary angioedema who had never had laryngeal edema.

3. The marker for predicting the onset site of hereditary angioedema type 2 according to claim 1, characterized in that: In patients with type 2 hereditary angioedema who had abdominal edema, the expression level of H3N3F1 was significantly higher than that in patients with type 2 hereditary angioedema who had never had abdominal edema; the expression levels of H6N3E1 and H5N2 were significantly lower than that in patients with type 2 hereditary angioedema who had never had abdominal edema.

4. Use of a marker for predicting the onset site of hereditary angioedema type 2 or a detection reagent thereof according to any one of claims 1 to 3 in the preparation of a product for predicting the onset site of hereditary angioedema type 2.

5. The use according to claim 4, characterized in that: The product is a medicine or a kit.

6. A product for predicting the onset site of hereditary angioedema type 2, characterized in that: The product comprises a marker for predicting the onset site of hereditary angioedema type 2 as described in any one of claims 1 to 3 or a detection reagent thereof.

7. The product according to claim 6, characterized in that The product is a medicine or a kit.

8. A device for predicting the onset site of hereditary angioedema type 2, characterized in that: The device comprises: A detection module, used for detecting the content of the marker for predicting the onset site of hereditary angioedema type 2 according to any one of claims 1 to 3 in a sample to be tested; An input module is used to obtain the detection result of the detection module; A judgment module, used for predicting the onset site of hereditary angioedema type 2 according to the test results obtained by the input module; Output module, used to output prediction results.

9. The device for predicting the onset site of hereditary angioedema type 2 according to claim 8, characterized in that: The judgment standard of the judgment module is: if the content of H6N5F1E2L1 and / or H5N4F1E1L1 in the sample to be tested is significantly higher than that in patients with type 2 hereditary angioedema who have never had laryngeal edema, then the edema onset site is predicted to be the larynx; If the content of H3N3F1 in the sample to be tested is significantly higher than that in patients with type 2 hereditary angioedema who have never experienced abdominal edema, and / or the expression levels of H6N3E1 and H5N2 are significantly lower than that in patients with type 2 hereditary angioedema who have never experienced abdominal edema, it is predicted that the site of edema onset is the abdomen.

10. The device for predicting the onset site of hereditary angioedema type 2 according to claim 8 or 9, characterized in that: The sample to be tested is plasma.

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