CN-121978348-A - Screening method for serum differential expression protein combination identified by NMOSD and MOGAD

Abstract

The present invention discloses a screening method for serum differentially expressed protein combinations used for distinguishing NMOSD from MOGAD, which relates to the field of screening technology for serum differentially expressed protein combinations. Multidimensional screening includes evaluating the expression stability of candidate differentially expressed proteins in the NMOSD patient group and MOGAD patient group. By removing proteins with large expression fluctuations or significant individual differences in the same disease population, the consistency of the retained proteins in the disease population is ensured. Compared with existing immune marker based diagnostic methods, the present invention analyzes the expression characteristics of coagulation related differentially expressed proteins in serum, which not only improves the discrimination rate between NMOSD and MOGAD, but also provides more accurate differentiation results in different stages of disease activity and remission by analyzing the expression characteristics of coagulation related differentially expressed proteins in serum.

Inventors

• SUN XIAOBO
• QIU WEI

Assignees

• 中山大学附属第三医院

Dates

Publication Date

20260505

Application Date

20260206

Claims (10)

1. 1. A screening method for the combination of NMOSD and MOGAD identified serum differential expression proteins is characterized by comprising the following steps: s1, respectively collecting peripheral blood samples of NMOSD patients and MOGAD patients, and separating to obtain serum samples; S2, carrying out protein expression detection on the serum sample to obtain serum protein expression data of NMOSD patients and MOGAD patients; S3, based on serum protein expression data, carrying out preliminary screening on protein expression differences between NMOSD patients and MOGAD patients to obtain a candidate differential expression protein set; S4, based on the expression stability, inter-disease difference amplitude and intra-group variation condition of the candidate differential expression proteins in two diseases, carrying out multidimensional screening on the candidate differential expression proteins, and removing proteins with insufficient distinguishing capacity or poor stability; s5, carrying out combined construction on the screened and reserved differential expression proteins, and forming candidate protein combinations by combining expression characteristics of a plurality of proteins; S6, evaluating the distinguishing capability of the candidate protein combination in NMOSD patients and MOGAD patients, and screening the protein combination with stable distinguishing effect between the two diseases; s7, combining proteins with stable distinguishing ability as serum differential expression proteins for NMOSD and MOGAD identification.
2. 2. The method according to claim 1, wherein the multidimensional screening in S4 comprises evaluating the expression stability of candidate differentially expressed proteins in the NMOSD patient group and MOGAD patient group by eliminating proteins with large expression fluctuation range or significant individual differences in the same disease population to ensure the consistency of the retained proteins in the disease population.
3. 3. A screening method for a combination of NMOSD and MOGAD-identified serum differentially expressed proteins according to claim 1, wherein the multidimensional screening in S4 further comprises evaluating the magnitude of the difference between the NMOSD and MOGAD of candidate differentially expressed proteins by preferentially retaining proteins whose expression differences between the two diseases are significant and whose direction of the difference is stable.
4. 4. A screening method for the combination of NMOSD and MOGAD-identified serum differentially expressed proteins according to claim 1, wherein the combined construction in S5 is not simply a superposition of a plurality of differentially expressed proteins, but is based on the complementarity of different proteins during disease differentiation, wherein complementarity includes at least complementarity of different proteins in terms of direction of expression change, magnitude of difference, or individual distribution characteristics.
5. 5. A screening method for differential expression of protein combinations in serum for NMOSD and MOGAD identification according to claim 1, wherein said evaluation of the discrimination ability of candidate protein combinations in step S6 comprises comparing the overall expression profile differences of different protein combinations in NMOSD patients and MOGAD patients and optimizing protein combinations based on the profile differences.
6. 6. A method of screening a combination of differentially expressed proteins in serum for use in the identification of NMOSD and MOGAD according to claim 1, wherein the combination of differentially expressed proteins in serum selected in S7 is made up of at least two differentially expressed proteins, and the composition of the combination of proteins is not dependent on the absolute expression level of a single protein, but rather is based on the relative expression profile of the proteins in the combination.
7. 7. The method according to claim 1, wherein the screening of candidate differentially expressed proteins in S4 is further based on calculation of the contribution of proteins to disease discrimination to quantify the discrimination ability of different proteins in NMOSD and MOGAD discrimination; The protein discrimination contribution is calculated according to the following formula: Wherein: represent the first A degree of differential contribution of the seed proteins; And (3) with Mean expression levels of the protein in NMOSD and MOGAD patients, respectively; And (3) with Respectively representing the expression discrete degree of the corresponding protein in two disease groups; Further, the degree of contribution of the discrimination is normalized, and the calculation method is as follows: By the calculation mode, proteins with higher contribution degrees are preferentially reserved and distinguished to enter the subsequent protein combination construction step.
8. 8. The method according to claim 1, wherein the step S6 is performed by constructing a protein combination discrimination score to evaluate the overall discrimination capability of different protein combinations between NMOSD and MOGAD; the protein combination discrimination score was calculated according to the following formula: Wherein: Representing a protein combination discrimination score; representing the first sample in the sample to be measured Expression level of the seed protein; Representing the average expression level of the corresponding protein in the reference disease population; A weight coefficient assigned for distinguishing the contribution degree according to the protein; Further, the protein combination discrimination scores of different disease populations were compared, and the discrimination index was calculated as follows: and carrying out quantitative evaluation on the disease discrimination capability of the protein combination through the discrimination score and the discrimination index.
9. 9. The method according to claim 1, wherein the evaluation of candidate protein combinations in S6 further comprises calculation of combination stability and robustness to evaluate the reliability of protein combinations under different sample conditions; The combined stability is calculated according to the following formula: Wherein: represents a protein combination stability score; And (3) with Respectively represent the first Average expression level and standard deviation of the seed proteins in the disease population; further, a combined robustness index is introduced, which is calculated as follows: Wherein: representing a protein combination robustness index; Is shown in the first Protein combination discrimination scores calculated in the secondary sample subset analysis; representing the calculated discrimination scores based on the population samples; protein combinations that still have stable discrimination ability under varying conditions of the sample are screened by a combination of stability scores and robustness indexes.
10. 10. The method for screening a combination of NMOSD and MOGAD identified serum differentially expressed proteins according to claim 1, wherein the method constructs a comprehensive identification decision index based on the multiple algorithm evaluation results for determining the final combination of proteins for NMOSD and MOGAD identification; The comprehensive identification decision index is calculated according to the following formula: Wherein: Representing the comprehensive identification decision index; a disease differentiation index representing a combination of proteins; representing a combined stability score; Representing a combined robustness index; Is a weight coefficient; further, when the comprehensive identification decision index satisfies the following condition, determining the protein combination as the target protein combination for the NMOSD and MOGAD identification: Through the comprehensive evaluation and decision rule, the system screening and optimization of serum differential expression protein combination are realized.

Description

Screening method for serum differential expression protein combination identified by NMOSD and MOGAD Technical Field The invention relates to the technical field of screening of serum differential expression protein combinations, in particular to a screening method for a serum differential expression protein combination identified by NMOSD and MOGAD. Background NMOSD and MOGAD are two different central nervous system inflammatory demyelinating diseases, although they overlap in clinical manifestations, however, there are significant differences in their pathogenesis, clinical progression and treatment methods, NMOSD is usually mediated by aquaporin 4 antibodies, while MOGAD is a disease caused by anti-MOG antibodies, since the clinical manifestations of these two diseases are often similar, especially during acute episodes, distinguishing them is of great importance for diagnosis and treatment, however, existing diagnostic methods rely mainly on antibody detection, such as anti-AQP 4 and anti-MOG antibodies, although these methods are widely used in clinic, their accuracy remains limited, especially at early stages of diagnosis. In addition, current research is mostly focused on the detection of immunological markers, but relatively few studies are conducted on the role of changes in the coagulation system in these diseases, which play a role not only in blood hemostasis and thrombosis but also are closely related to immune responses, and in some immune-related diseases, abnormalities of the coagulation system are considered as important factors for the occurrence and development of diseases, in particular, the expression changes of coagulation-related proteins, possibly becoming potential biomarkers for diagnosing and distinguishing between different immune diseases; Therefore, how to help distinguish between NMOSD and MOGAD by the expression characteristics of coagulation-related differential proteins remains an important topic in current research; Although the coagulation system has shown potential diagnostic value in some immune diseases, there is no existing systematic technical solution for how to effectively screen and evaluate coagulation related proteins, especially in differential diagnosis of NMOSD and MOGAD, and therefore, there is a need to develop a new method to further improve the diagnostic accuracy and discrimination ability of these two diseases by combining proteins differentially expressed by serum. Disclosure of Invention The invention provides a screening method for serum differential expression protein combination identified by NMOSD and MOGAD, which can effectively solve the problems in the background technology. In order to achieve the aim, the invention provides the following technical scheme that the screening method for the combination of NMOSD and MOGAD identified serum differential expression proteins comprises the following steps: s1, respectively collecting peripheral blood samples of NMOSD patients and MOGAD patients, and separating to obtain serum samples; S2, carrying out protein expression detection on the serum sample to obtain serum protein expression data of NMOSD patients and MOGAD patients; S3, based on serum protein expression data, carrying out preliminary screening on protein expression differences between NMOSD patients and MOGAD patients to obtain a candidate differential expression protein set; S4, based on the expression stability, inter-disease difference amplitude and intra-group variation condition of the candidate differential expression proteins in two diseases, carrying out multidimensional screening on the candidate differential expression proteins, and removing proteins with insufficient distinguishing capacity or poor stability; s5, carrying out combined construction on the screened and reserved differential expression proteins, and forming candidate protein combinations by combining expression characteristics of a plurality of proteins; S6, evaluating the distinguishing capability of the candidate protein combination in NMOSD patients and MOGAD patients, and screening the protein combination with stable distinguishing effect between the two diseases; s7, combining proteins with stable distinguishing ability as serum differential expression proteins for NMOSD and MOGAD identification. According to the technical scheme, the multidimensional screening in the S4 comprises evaluation of the expression stability of the candidate differential expression proteins in the NMOSD patient group and the MOGAD patient group, and the consistency of the reserved proteins in the disease group is ensured by eliminating the proteins with larger expression fluctuation range or obvious individual difference in the same disease group. According to the above technical solution, the multidimensional screening in S4 further comprises evaluating the difference amplitude between NMOSD and MOGAD of the candidate differentially expressed proteins, by preferentially retaining the