Method for analyzing serum differential expression protein characteristics of AQP4-IgG positive NMOSD patient

Abstract

The invention discloses a method for analyzing serum differential expression protein characteristics of an AQP4-IgG positive NMOSD patient, and relates to the technical field of differential expression protein characteristic analysis, and the method comprises the following steps: S1, collecting a serum sample of a to-be-detected subject; s2, carrying out protein detection on the serum sample to obtain expression level data of at least one or more of PRDX2, CLU, ECM1, CFD, GPI and S100A8, wherein the expression level data is one or more of PRDX2, CLU, ECM1, CFD, GPI and S100A8; s3, carrying out comparative analysis on the expression level of the serum protein and a pre-established AQP4-IgG positive NMOSD serum protein expression characteristic reference standard; according to the method for auxiliary diagnosis of the AQP4-IgG positive neuromyelitis optica pedigree disease provided by the invention, a judgment mode capable of reflecting molecular characteristics of the AQP4-IgG positive neuromyelitis optica pedigree disease is constructed by carrying out conjoint analysis on various serum protein expression characteristics related to immunoregulation, inflammatory response, complement activation and metabolism; therefore, the problem of insufficient detection stability of a single biomarker is avoided.

Description

Technical Field

[0001] This invention relates to the field of differentially expressed protein characterization technology, specifically a method for analyzing the differentially expressed protein characterization in the serum of AQP4-IgG positive NMOSD patients. Background Technology

[0002] Neuromyelitis Optica Spectrum Disorders (NMOSD) is a group of inflammatory demyelinating diseases of the central nervous system mediated by autoimmune abnormalities. Clinically, it is mainly characterized by optic neuritis and longitudinal extensive transverse myelitis. The disease progresses rapidly and has a high risk of disability. Studies have shown that aquaporin-4 (AQP4) autoantibodies play an important role in the pathogenesis of NMOSD, and serum AQP4-IgG positivity has become one of the important criteria for the diagnosis of NMOSD. However, because NMOSD has significant overlap with other central nervous system demyelinating diseases in terms of clinical manifestations, imaging features and disease progression, and there are large individual differences among patients, relying solely on single antibody detection or imaging examinations is difficult to fully reflect the molecular characteristics and pathological state of the disease, thus still having certain limitations in clinical diagnosis, differential diagnosis and disease assessment.

[0003] In existing research, serum-based proteomics analysis is widely used to explore the potential molecular mechanisms and candidate biomarkers of NMOSD. However, most of these studies focus on the screening of differentially expressed proteins and the analysis of mechanisms, remaining primarily at the research level. There is a lack of systematic methodological design tailored to clinical applications. On the one hand, single serum protein indicators are easily affected by individual differences, detection conditions, and disease heterogeneity, resulting in limited stability and reproducibility. On the other hand, current technologies lack a clear diagnostic logic based on the combined expression characteristics of multiple serum proteins, making it difficult to provide operable auxiliary diagnostic criteria for clinical use. Therefore, there is an urgent need for an AQP4-IgG positive NMOSD auxiliary diagnostic method that can comprehensively analyze multiple serum protein expression characteristics and has clear diagnostic criteria to address the accuracy and practicality issues of existing technologies. Summary of the Invention

[0004] This invention provides an analytical method for the differentially expressed protein characteristics in the serum of AQP4-IgG positive NMOSD patients, which can effectively solve the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients, comprising the following steps: S1. Collect serum samples from the subjects to be tested; S2. Perform protein detection on serum samples to obtain expression level data of at least the following serum proteins, including at least PRDX2, CFD, and S100A8. S3. Compare and analyze the expression levels of serum proteins with the pre-established reference standard for the expression characteristics of AQP4-IgG positive NMOSD serum proteins; S4. When the expression pattern of serum proteins meets the reference standard, the subject is determined to have the risk of AQP4-IgG positive neuromyelitis optica spectrum disorder or to be diagnosed as AQP4-IgG positive NMOS.

[0006] According to the above technical solution, the serum protein expression characteristics are composed of a variety of serum proteins related to immune regulation, complement activation, oxidative stress response, extracellular matrix remodeling or energy metabolism. The serum proteins include at least three or more of PRDX2, CLU, ECM1, CFD, GPI, and S100A8.

[0007] According to the above technical solution, the expression level of the serum protein is obtained by detecting the concentration, relative abundance, signal intensity, or combination thereof of the target protein in the serum sample of the subject to be tested, and the detection results can be characterized by absolute quantification or relative quantification.

[0008] According to the above technical solution, the comparative analysis includes a joint analysis of the expression levels of at least three or more serum proteins. The joint analysis is not based on the independent determination of a single protein, but rather on the comprehensive determination result that can reflect the characteristics of AQP4-IgG positive neuromyelitis optica spectrum disorder by comprehensively considering the synergistic changes of multiple serum proteins in immune regulation, inflammatory response, complement activation, oxidative stress or metabolic-related pathways.

[0009] According to the above technical solution, the method is further used to distinguish patients with AQP4-IgG positive neuromyelitis optica spectrum disorder from patients with myelin oligodendrocyte glycoprotein antibody-related diseases or healthy individuals. Specifically, by comparing the difference patterns of serum protein expression characteristics under different disease states, protein expression combinations with characteristics of AQP4-IgG positive neuromyelitis optica spectrum disorder are identified.

[0010] According to the above technical solution, the reference standard for serum protein expression characteristics of AQP4-IgG positive neuromyelitis optica spectrum disease was established by statistical analysis of serum samples from multiple patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disease. The reference standard includes the expression range, expression trend, or combined expression pattern of at least one serum protein.

[0011] According to the above technical solution, the detection methods for serum proteins include immunological detection methods, antibody- or antigen-based detection methods, mass spectrometry detection methods, or any combination of the above detection methods.

[0012] According to the above technical solution, the serum protein expression characteristics further include at least one reference interval or trend rule for determining abnormal expression, wherein the reference interval or trend rule is established based on the statistical analysis results of healthy control populations and patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disorder.

[0013] According to the above technical solution, the serum protein expression characteristics are further used to assess the disease activity status, relapse risk, or disease progression trend of patients with AQP4-IgG positive neuromyelitis optica spectrum disorders. Specifically, the changes in serum protein expression levels over time are analyzed by detecting and comparing serum samples from the same subject collected at different time points.

[0014] According to the above technical solution, the process of obtaining, comparing, analyzing and determining the serum protein expression level can be completed with the assistance of a data processing system or a computer program. The data processing system includes an input module for receiving serum protein detection data, a storage module for storing serum protein expression characteristic reference standards, and a data processing module for executing comparison analysis and determination logic.

[0015] Compared with existing technologies, the beneficial effects of this invention are as follows: This invention has a scientifically sound and reasonable structure, is safe and convenient to use, and provides a method for assisting in the diagnosis of AQP4-IgG positive neuromyelitis optica spectrum disorders. This method constructs a judgment model that reflects the molecular characteristics of AQP4-IgG positive neuromyelitis optica spectrum disorders by jointly analyzing the expression characteristics of various serum proteins related to immune regulation, inflammatory response, complement activation, and metabolism. This avoids the problem of insufficient stability in single biomarker detection. The method described in this invention has a clear judgment logic and good reproducibility, which can improve the accuracy and consistency of auxiliary diagnosis of diseases and help differentiate NMOSD from other central nervous system demyelinating diseases. It provides a reliable auxiliary basis for clinical diagnostic decisions, disease assessment, and subsequent treatment selection, and has good clinical application prospects. Attached Figure Description

[0016] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof.

[0017] In the attached diagram: Figure 1 This is a schematic diagram of the overall process of the serum protein characteristic-assisted diagnostic method of the present invention; Figure 2 This is a schematic diagram of the multi-protein joint determination logic of the present invention. Detailed Implementation

[0018] The preferred embodiments of the present invention will be described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are for illustration and explanation only and are not intended to limit the present invention.

[0019] Example: Figure 1-2 As shown, the present invention provides a technical solution: an analytical method for the differentially expressed protein characteristics in the serum of AQP4-IgG positive NMOSD patients, comprising the following steps: S1. Collect serum samples from the subjects to be tested; S2. Perform protein detection on serum samples to obtain expression level data of at least the following serum proteins, including at least PRDX2, CFD, and S100A8. S3. Compare and analyze the expression levels of serum proteins with the pre-established reference standard for the expression characteristics of AQP4-IgG positive NMOSD serum proteins; S4. When the expression pattern of serum proteins meets the reference standard, the subject is determined to have the risk of AQP4-IgG positive neuromyelitis optica spectrum disorder or to be diagnosed as AQP4-IgG positive NMOS.

[0020] According to the above technical solution, the serum protein expression characteristics are composed of a variety of serum proteins related to immune regulation, complement activation, oxidative stress response, extracellular matrix remodeling or energy metabolism. Serum proteins include at least three or more of PRDX2, CLU, ECM1, CFD, GPI and S100A8; Among them, PRDX2 is used to reflect abnormal changes in oxidative stress-related pathways, CLU is used to reflect the status of complement regulation and cell protection function, ECM1 is used to reflect extracellular matrix-related pathological changes, CFD is used to reflect the level of complement bypass activation, GPI is used to reflect the status of glucose metabolism-related pathways, and S100A8 is used to reflect the degree of inflammatory response and innate immune activation. By jointly analyzing the expression levels of the above-mentioned multiple serum proteins, a serum protein expression characteristic pattern for AQP4-IgG positive neuromyelitis optica spectrum disease was constructed, thereby improving the accuracy of auxiliary diagnosis of the disease and reducing the risk of misjudgment caused by single biomarker detection.

[0021] According to the above technical solution, the expression level of serum protein is obtained by detecting the concentration, relative abundance, signal intensity or combination thereof of the target protein in the serum sample of the subject to be tested. The detection results can be characterized by absolute quantification or relative quantification. The relative quantification method includes comparing the detection results of the target protein in the serum of the subjects to be tested with the expression levels of the corresponding protein in healthy controls, patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disorder, or a pre-established reference database. Expression levels can be expressed as protein concentration values, relative expression folds, standardized numerical ranges, or expression trends, and serve as the basic data for constructing serum protein expression characteristic patterns, thereby reflecting changes in the pathological state of AQP4-IgG positive neuromyelitis optica spectrum disorders in the subjects being tested.

[0022] According to the above technical solution, the comparative analysis includes the joint analysis of the expression levels of at least three or more serum proteins. The joint analysis is not based on the independent determination of a single protein, but rather on the comprehensive consideration of the synergistic changes of multiple serum proteins in immune regulation, inflammatory response, complement activation, oxidative stress or metabolic-related pathways to construct a comprehensive judgment result that can reflect the characteristics of AQP4-IgG positive neuromyelitis optica spectrum disease. The combined analysis method includes, but is not limited to, weighted calculation, logical combination judgment, graded scoring or pattern matching analysis of the expression levels of multiple serum proteins. When the expression changes of multiple serum proteins meet the preset serum protein expression characteristic pattern, a comprehensive judgment conclusion is output to assist in the diagnosis of AQP4-IgG positive neuromyelitis optica spectrum disease, thereby improving the stability and reliability of the diagnostic results.

[0023] According to the above technical solution, the method is further used to distinguish patients with AQP4-IgG positive neuromyelitis optica spectrum disease from patients with myelin oligodendrocyte glycoprotein antibody-related diseases or healthy individuals. In particular, by comparing the difference patterns of serum protein expression characteristics under different disease states, protein expression combinations with AQP4-IgG positive neuromyelitis optica spectrum disease characteristics are identified. Differentiation methods include determining whether there are significant differences in the expression direction, variation range, or combination characteristics of serum proteins in different disease populations, thereby providing auxiliary basis for differential diagnosis, disease classification, or selection of clinical treatment plans, and reducing the risk of misdiagnosis or missed diagnosis due to similar clinical manifestations.

[0024] According to the above technical solution, the reference standard for serum protein expression characteristics of AQP4-IgG positive neuromyelitis optica spectrum disease was established by statistical analysis of serum samples from multiple patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disease. The reference standard includes the expression range, expression trend, or combined expression pattern of at least one serum protein. Reference standards can be stored in the form of databases, feature templates, or judgment rules, and can be called when analyzing subjects to be tested, so as to realize automatic or semi-automatic comparison of serum protein expression characteristics, thereby improving the reproducibility, consistency and clinical feasibility of the method.

[0025] According to the above technical solution, the detection methods for serum proteins include immunological detection methods, antibody or antigen-based detection methods, mass spectrometry detection methods, or any combination of the above detection methods. The detection methods are applicable to clinical sample testing, medical testing, or scientific research analysis scenarios. The method can be applied in hospital laboratories, medical testing laboratories, or research institutions to assist in the diagnosis, risk assessment, or disease subtyping analysis of patients suspected of having AQP4-IgG positive neuromyelitis optica spectrum disorders, thereby expanding the application scope of the method in clinical practice and scientific research translation.

[0026] According to the above technical solution, the serum protein expression characteristics further include at least one reference interval or trend rule for determining abnormal expression. The reference interval or trend rule is established based on the statistical analysis results of healthy control groups and patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disease. The reference range can be the expression range of a single serum protein or a combination range of expression levels of multiple serum proteins. The trend rules include the expression pattern of the target serum protein showing continuous upregulation, continuous downregulation, or synergistic changes relative to healthy control samples. When the serum protein expression level of the subject to be tested falls within the reference range or meets the trend rules, it is determined that its serum protein expression characteristics are related to AQP4-IgG positive neuromyelitis optica spectrum disease, thereby improving the objectivity and consistency of the determination process and reducing the influence of subjective judgment factors on the diagnostic results.

[0027] According to the above technical solution, serum protein expression characteristics are further used to assess the disease activity status, relapse risk or disease progression trend of patients with AQP4-IgG positive neuromyelitis optica spectrum disorders. Specifically, by detecting and comparing serum samples of the same subject collected at different time points, the changes in serum protein expression levels over time are analyzed. When the expression level of serum protein shows an increasing, expanding, or abnormally aggravated trend in continuous monitoring, it is determined that the subject may have an increased risk of disease activity or relapse. The assessment method can provide clinicians with auxiliary reference information during disease follow-up, treatment plan adjustment or efficacy evaluation, thereby expanding the application value of the method in disease management and prognosis assessment.

[0028] According to the above technical solution, the process of obtaining, comparing, analyzing and determining serum protein expression levels can be completed by a data processing system or computer program. The data processing system includes an input module for receiving serum protein detection data, a storage module for storing serum protein expression characteristic reference standards, and a data processing module for executing comparison analysis and determination logic. The data processing module is used to compare and analyze the serum protein expression data of the subjects to be tested with the pre-established serum protein expression characteristics of AQP4-IgG positive neuromyelitis optica spectrum disease, and output the determination results for auxiliary diagnosis, risk assessment or disease classification. By introducing a systematic data processing approach, the efficiency and consistency of the method in large-scale sample testing can be improved, and its feasibility in clinical testing and scientific research applications can be enhanced.

[0029] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients, characterized in that: Includes the following steps: S1. Collect serum samples from the subjects to be tested; S2. Perform protein detection on serum samples to obtain expression level data for at least one or more of the following serum proteins: PRDX2, CLU, ECM1, CFD, GPI, and S100A8; S3. Compare and analyze the expression levels of serum proteins with the pre-established reference standard for the expression characteristics of AQP4-IgG positive NMOSD serum proteins; S4. When the expression pattern of serum proteins meets the reference standard, the subject is determined to have the risk of AQP4-IgG positive neuromyelitis optica spectrum disorder or to be diagnosed as AQP4-IgG positive NMOS.

2. The method for analyzing the differentially expressed protein characteristics in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The serum protein expression characteristics are composed of a variety of serum proteins that are related to immune regulation, complement activation, oxidative stress response, extracellular matrix remodeling or energy metabolism. The serum proteins include at least three or more of PRDX2, CLU, ECM1, CFD, GPI, and S100A8.

3. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The expression level of the serum protein is obtained by detecting the concentration, relative abundance, signal intensity, or a combination thereof of the target protein in the serum sample of the subject to be tested. The detection results can be characterized by absolute quantification or relative quantification.

4. The method for analyzing the differentially expressed protein characteristics in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The comparative analysis includes a combined analysis of the expression levels of at least three or more serum proteins. This combined analysis is not based on the independent determination of a single protein, but rather on the comprehensive assessment of the synergistic changes of multiple serum proteins in immune regulation, inflammatory response, complement activation, oxidative stress, or metabolic-related pathways, to construct a comprehensive assessment result that reflects the characteristics of AQP4-IgG positive neuromyelitis optica spectrum disorders.

5. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The method is further used to distinguish patients with AQP4-IgG positive neuromyelitis optica spectrum disorder from patients with myelin oligodendrocyte glycoprotein antibody-associated disease or healthy individuals. Specifically, it identifies protein expression combinations characteristic of AQP4-IgG positive neuromyelitis optica spectrum disorder by comparing the differential patterns of serum protein expression features in different disease states.

6. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The reference standard for serum protein expression characteristics of AQP4-IgG positive neuromyelitis optica spectrum disorder was established through statistical analysis of serum samples from multiple patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disorder. The reference standard includes the expression range, expression trend, or combined expression pattern of at least one serum protein.

7. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The detection methods for serum proteins include immunological detection methods, antibody- or antigen-based detection methods, mass spectrometry detection methods, or any combination of the above methods.

8. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The serum protein expression characteristics further include at least one reference interval or trend rule for determining abnormal expression, which is established based on statistical analysis results of healthy controls and patients diagnosed with AQP4-IgG positive neuromyelitis optica spectrum disorder.

9. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The serum protein expression characteristics are further used to assess the disease activity status, relapse risk, or disease progression trend of patients with AQP4-IgG positive neuromyelitis optica spectrum disorders. Specifically, the changes in serum protein expression levels over time are analyzed by detecting and comparing serum samples from the same subject collected at different time points.

10. The method for analyzing the characteristics of differentially expressed proteins in the serum of AQP4-IgG positive NMOSD patients according to claim 1, characterized in that, The process of acquiring, comparing, analyzing, and determining the expression level of serum proteins can be completed with the assistance of a data processing system or a computer program. The data processing system includes an input module for receiving serum protein detection data, a storage module for storing reference standards for serum protein expression characteristics, and a data processing module for executing comparative analysis and determination logic.

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