Biomarker for auxiliary diagnosis of progressive suprakaryotic paralysis and application thereof

Abstract

The invention discloses a biomarker for auxiliary diagnosis of progressive suprakaryotic paralysis and application thereof, and relates to the technical field of biomedicine. The biomarker is a neuron-derived extracellular vesicle carrying a Tau deformable protein; the Tau deformation protein is 4R Tau or pTau181, and the Tau deformation protein is a protein with the molecular weight of 4R The invention develops a biomarker capable of assisting in diagnosing progressive suprakaryotic paralysis, and the biomarker can be used for effectively identifying a progressive suprakaryotic paralysis (PSP) patient, a Parkinson's disease (PD) patient and a healthy person (HC). Meanwhile, a progressive supranuclear paralysis diagnosis model is also constructed, the sensitivity of the multivariable logistic regression model for distinguishing PSP and HC is 95.7%, the specificity is 90.0%, and the AUC is 0.974; the sensitivity of the multivariable logistic regression model for distinguishing PSP and PD is 96.3%, the specificity is 90.0%, and the AUC is 0.968.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to biomarkers for assisting diagnosis of progressive supranuclear palsy and applications thereof. Background Art

[0002] Progressive supranuclear palsy (PSP) is a common atypical Parkinson's syndrome, which is pathologically characterized by abnormal aggregation of Tau protein in neurons, oligodendrocytes and astrocytes. Since Parkinson's disease (PD) and other atypical parkinsonian syndromes have multiple clinical subtypes and symptom overlap, the diagnosis is very complicated. This diagnostic complexity often leads to a high misdiagnosis rate. Reliable biomarkers are urgently needed to facilitate early diagnosis and distinguish PSP from other neurodegenerative diseases.

[0003] Existing studies on the detection of Tau protein: Abnormal Tau protein deposition has been detected in some biological fluids and peripheral tissues (including plasma, olfactory and oral epithelial cells, colon specimens, and skin biopsies). Compared with tissue biopsy and other methods of acquisition, plasma is a clinical sample that can be routinely obtained.

[0004] In the current study, disease-related extracellular vesicles (EVs) were extracted and analyzed from plasma by immunocapture and enzyme-linked immunosorbent assay (ELISA) methods, especially focusing on Tau protein and its phosphorylated form. These methods require a large volume of serum sample (usually 500 μL), and the detection process is relatively complicated and cumbersome.

[0005] Although the prior art has made some progress in the study of biomarkers for neurodegenerative diseases, the following problems and defects still exist: high sample volume requirement: the prior art usually requires 500 μL or more serum samples; complex detection process: the existing immunocapture and ELISA detection steps are cumbersome and time-consuming, requiring multiple operations, which may lead to errors between steps, thereby affecting the reliability and repeatability of the results; insufficient sensitivity and specificity; lack of non-invasive detection methods: existing biological samples need to be obtained through invasive operations such as tissue biopsy, which is difficult to be widely used in conventional medical treatment. The present invention hopes to provide an effective and non-invasive detection method for the early diagnosis of PSP, so as to effectively distinguish PSP from other neurodegenerative diseases. Summary of the invention

[0006] The purpose of the present invention is to provide a biomarker for assisting the diagnosis of progressive supranuclear palsy and its application, so as to solve the problems existing in the above-mentioned prior art. The present invention provides a new biomarker and a diagnostic model for assisting the diagnosis of progressive supranuclear palsy, especially for distinguishing progressive supranuclear palsy from Parkinson's disease, and has important clinical application value.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides a biomarker for assisting the diagnosis of progressive supranuclear palsy, wherein the biomarker is a neuron-derived extracellular vesicle carrying Tau deformed protein;

[0009] The Tau deformed protein is 4R Tau or pTau181.

[0010] Furthermore, a higher level of the biomarker in plasma indicates that the patient has a higher risk of developing progressive supranuclear palsy.

[0011] The present invention also provides a biomarker for distinguishing progressive supranuclear palsy and Parkinson's disease, wherein the biomarker is a neuron-derived extracellular vesicle carrying Tau deformed protein;

[0012] The Tau deformed protein is 4R Tau or pTau181.

[0013] Furthermore, a higher content of the biomarker in plasma indicates that the patient has a higher risk of developing progressive supranuclear palsy among progressive supranuclear palsy and Parkinson's disease.

[0014] The present invention also provides a diagnostic model ND Tau / Tau for progressive supranuclear palsy, wherein the diagnostic model NDTau / Tau uses the plasma concentration of neuron-derived extracellular vesicles carrying Tau protein and the plasma concentration of total extracellular vesicles carrying Tau protein as input variables;

[0015] The ratio diagnostic model is: risk value R = plasma concentration of neuron-derived extracellular vesicles carrying Tau protein / plasma concentration of total extracellular vesicles carrying Tau protein;

[0016] A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy.

[0017] The present invention also provides a diagnostic model ND pTau231 / pTau231 for progressive supranuclear palsy, wherein the diagnostic model ND pTau231 / pTau231 uses the plasma concentration of neuron-derived extracellular vesicles carrying pTau231 protein and the plasma concentration of total extracellular vesicles carrying pTau231 protein as input variables;

[0018] The ratio diagnostic model is risk value R=plasma concentration of neuron-derived extracellular vesicles carrying pTau231 protein / plasma concentration of total extracellular vesicles carrying pTau231 protein;

[0019] A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy.

[0020] The present invention also provides a diagnostic model ND 4R Tau / 4R Tau for distinguishing progressive supranuclear palsy from Parkinson's disease, wherein the diagnostic model ND 4R Tau / 4R Tau uses the plasma concentration of neuron-derived extracellular vesicles carrying 4R Tau protein and the plasma concentration of total extracellular vesicles carrying 4R Tau protein as input variables;

[0021] The ratio diagnostic model is: risk value R = plasma concentration of neuron-derived extracellular vesicles carrying 4R Tau protein / plasma concentration of total extracellular vesicles carrying 4R Tau protein;

[0022] A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy among progressive supranuclear palsy and Parkinson's disease.

[0023] The present invention also provides a multivariate logistic regression model for assisting in the diagnosis of progressive supranuclear palsy. When used to distinguish patients with progressive supranuclear palsy from normal persons, the risk value D1 of the multivariate logistic regression model is D1=0.0000054431×C tau +0.0000043461×C 4R Tau +0.0000004477×C pTau181 +0.0000032264×C pTau231 -0.0000001555×C pTau396 ;

[0024] When used to distinguish patients with progressive supranuclear palsy from patients with Parkinson's disease, the risk value of the multivariate logistic regression model was D2 = 0.0000070751 × C tau +0.0000028567×C 4RTau +0.0000006862×C pTau181 +0.0000013800×C pTau231 -0.0000004485×C pTau396 ;

[0025] The C tau , said C 4RTau , said C pTau181 , said C pTau231 and the C pTau396 Represents the concentration of neuronal-derived extracellular vesicles containing Tau, 4R Tau, pTau181, pTau231 or pTau396 in plasma, respectively.

[0026] The present invention also provides the use of a reagent for detecting the content of the above-mentioned biomarker in plasma in the preparation of a kit for auxiliary diagnosis of progressive supranuclear palsy.

[0027] Furthermore, the reagent is a reagent for nanoscale flow cytometry detection.

[0028] The present invention discloses the following technical effects:

[0029] The present invention develops a biomarker that can assist in the diagnosis of progressive supranuclear palsy, which can effectively distinguish patients with progressive supranuclear palsy (PSP), patients with Parkinson's disease (PD) and healthy people (HC). At the same time, a diagnostic model for progressive supranuclear palsy is also constructed, wherein the sensitivity of the multivariate logistic regression model for distinguishing PSP from HC is 95.7%, the specificity is 90.0%, and the AUC is 0.974; the sensitivity of the multivariate logistic regression model for distinguishing PSP from PD is 96.3%, the specificity is 90.0%, and the AUC is 0.968.

[0030] The present invention provides a new biomarker and a diagnostic model for assisting the diagnosis of progressive supranuclear palsy, especially for distinguishing progressive supranuclear palsy from Parkinson's disease, and has important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only 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.

[0032] Figure 1 are the characteristic analysis results of EVs; A is the TEM image of EVs; B is the Western blot analysis of ultracentrifugation supernatant (UCSupernatant), ultracentrifuged EVs and untreated reference plasma samples (Raw Plasma); C is the distribution map of nanoparticles in EVs of the PSP group; D is the distribution map of nanoparticles in EVs of the PD group; E is the distribution map of nanoparticles in EVs of the HCs group; F is a statistical graph of the total concentration of EVs of all particle sizes in the plasma of the PSP, PD and HCs groups;

[0033] Figure 2The results of concentration detection and diagnostic value analysis of neuronal EVs carrying Tau or its deformed proteins in plasma samples of different groups; AE are statistical graphs of the concentrations of neuronal EVs containing Tau, 4R Tau, pTau181, pTau231 and pTau396 in the plasma of PSP, PD and HCs, respectively; FJ are statistical graphs of the ratio of neuronal EVs carrying Tau or its deformed proteins to the total amount of EVs carrying Tau or its deformed proteins in the plasma of PSP, PD and HC patients, respectively; K is the receiver operating characteristic curve of neuronal EVs carrying Tau or its deformed proteins in plasma for distinguishing between PSP and HC patients; L is the receiver operating characteristic curve of the multivariate logistic regression model for distinguishing between PSP and HC patients; M is the receiver operating characteristic curve of neuronal EVs carrying Tau or its deformed proteins in plasma for distinguishing between PSP and PD patients; N is the receiver operating characteristic curve of the multivariate logistic regression model for distinguishing between PSP and PD patients. DETAILED DESCRIPTION

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0037] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0038] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0039] Terminology Note:

[0040] Tau protein is a microtubule-associated protein that is widely distributed in the nerve cells of the nervous system and is an important component used to stabilize the microtubules that serve as the skeleton of nerve cells.

[0041] 4R tau refers to Tau protein with 4 repeat regions.

[0042] pTau181, pTau231 and pTau396 are different phosphorylated forms of Tau protein, namely phosphorylated Tau-181 protein, phosphorylated Tau-231 protein and phosphorylated Tau-396 protein.

[0043] Example 1

[0044] 1. Materials and Methods

[0045] Included samples: 30 PSP patients, 27 PD patients and 23 healthy controls (HC).

[0046] Preparation of plasma samples: Venous blood samples were collected from fasting participants in the morning using test tubes coated with ethylenediaminetetraacetic acid. The blood samples were then centrifuged at 1500×g for 15 minutes (4°C) to obtain the first supernatant, and the first supernatant was centrifuged at 12,000×g for 30 minutes (4°C) to obtain the second supernatant, which was the plasma sample. The second supernatant was stored at -80°C for nanoscale flow cytometry and nanoparticle tracking analysis (NTA).

[0047] Isolation of neuronal EVs: EVs carrying NMDAR2A were isolated from ultrapure plasma EVs using an immunocapture protocol. 10 μg of anti-NMDAR2A antibody was coated on 1 mg of M-270 epoxy beads using the Dynabeads Antibody Coupling Kit (14311D, Invitrogen) according to the manufacturer's instructions.

[0048] Transmission electron microscopy (TEM) analysis: 5 μL of NMRAR2A-labeled EVs were placed on a copper grid coated with a carbon support film. After the sample was incubated for 60 seconds, the excess sample was removed with filter paper. The copper grid was then stained with a 2% uranyl acetate solution for 60 seconds. The excess uranyl acetate solution was removed with filter paper, and the sample was then air-dried. Electron microscopy images were taken using a Tecnai F20 transmission electron microscope at 200 kV.

[0049] Western blot analysis: The total protein concentration in ultracentrifugation supernatants, ultracentrifuged EVs, and untreated reference plasma samples was assessed using the BCA method. 15 μg of total protein contained in each sample was separated using 4-20% Bis-Tris gel (M42010C, Genscript) and then electrotransferred to nitrocellulose membranes. After blocking with 5% skim milk, the membranes were incubated with anti-NMDAR2A antibody, Ali polyclonal antibody, and CD9 monoclonal antibody overnight at 4°C.

[0050] Nanoparticle tracking analysis: 2 μL of plasma samples from PSP patients, PD patients, and HCs were diluted 1:500 in PBS (pH 7.4). These diluted samples were then analyzed using the ZetaView platform (Particle Metrix) to assess the distribution and concentration of nanoparticles.

[0051] Analysis of EVs using Cytoflex nano-flow cytometer: Antibodies were conjugated to fluorophores using the Zenon IgG labeling kit according to the manufacturer’s instructions. Antibodies against NMDAR2A (neuronal origin) and Tau and its different phosphorylated forms (e.g., pTau181, pTau231, pTau396) were labeled using different fluorescent labeling kits. 5 μL of plasma was placed in a flow cytometer tube. Then 0.1 μg of fluorescently conjugated NMDAR2A antibody was added and incubated for 30 min, followed by 0.2 μg of fluorescently conjugated Tau antibody and incubated for 20 min, all at room temperature and in the dark. After incubation, the mixture was diluted 1:60 with PBS, vortexed, and centrifuged for 10 seconds. Vesicles below 500 nm were quantitatively detected using a nano-flow cytometer on a Cytoflex S platform (Beckman Coulter, Milano, Italy) in VSSC-H mode. The concentration of EVs carrying Tau protein and NMDAR2A was calculated according to the flow rate and PBS dilution ratio. All samples were tested within 4 hours and a single batch of cell counting analysis was performed on the same day.

[0052] The present invention utilizes nanoscale flow cytometry to rapidly and sensitively analyze neuron-derived EVs in plasma, and achieves the technical effect of improving the early diagnosis capability of progressive supranuclear palsy by combining fluorescent labeling of specific antibodies and an efficient sample processing process.

[0053] 2. Data Processing

[0054] Statistical analysis was performed using GraphPad Prism 10 and SPSS 26.0. The Kruskal-Wallis test was used to assess the intergroup differences in the mean concentration of neuronal EVs carrying Tau, and the Dunn test was then used to compare PSP with PD and PSP with HC. Receiver operating characteristic (ROC) curve analysis was used to assess the sensitivity and specificity of these markers in distinguishing PSP from HCs or PD. A multivariate model was established using binary logistic regression, in which multiple neuronal EVs carrying Tau protein were included to distinguish PSP from PD or HC. Spearman correlation analysis was used to assess the correlation between clinical indicators and the concentration of neuronal EVs carrying Tau in the PSP group, and Bonferroni correction was used to adjust for multiple comparisons. All tests were two-tailed, and statistical significance was set at P < 0.05.

[0055] 3. Results and Analysis

[0056] 1. Characteristics of EVs

[0057] TEM analysis showed that the diameter of neuronal EVs was approximately 100 nm ( Figure 1 Western blot analysis showed that NMDAR2A was enriched together with the general EV markers Alix and CD9 in reference EVs samples obtained by ultracentrifugation ( Figure 1 NTA analysis showed that a broad peak centered at 100 nm appeared in the PD, PSP, and HC groups ( Figure 1 There was no significant difference in EV concentrations between these groups ( Figure 1 Middle F).

[0058] 2. Neuronal EVs carrying Tau can effectively distinguish PSP from PD and HC

[0059] Among the 30 PSP, 27 PD, and 23 HC included, the concentrations of neuronal EVs containing Tau, 4R Tau, and pTau181 in the plasma of PSP patients were significantly higher than those of HC and PD patients (PSP vs. HCs: P Tau <0.0001, P 4RTau =0.004, P pTau181 <0.001; PSP vs PD: P Tau <0.0001, P 4RTau <0.001, P pTau181 =0.005; Kruskal-Wallis test and Dunn's multiple comparison test, Figure 2 (in AC).

[0060] The present invention also analyzed the ratio of neuron-derived EVs carrying Tau, 4R Tau and pTau181, pTau231, and pTau396 to the total EVs carrying Tau (referred to as ratio). Figure 2 FJ. The ratio of neuronal EVs carrying Tau to EVs carrying Tau in PSP was significantly higher than that in PD (P = 0.001) and HCs (P < 0.0001) ( Figure 2 In addition, the ratio of neuronal EVs carrying 4RTau to the total number of EVs carrying 4R Tau was higher in PSP than in PD (P = 0.003) ( Figure 2 Middle G), the ratio of neuron-derived EVs carrying pTau231 to the total number of EVs carrying pTau231 was higher in PSPs than in HCs (P = 0.007) ( Figure 2 Middle I).

[0061] ROC analysis evaluated the diagnostic utility of neuronal EVs containing Tau, 4R Tau, pTau181, pTau231, and pTau396 in plasma in differentiating PSP from HC. The cut-off values ​​(unit: cells / mL) of these markers were as follows: Tau, 697500; 4R Tau, 1327500; pTau181, 3544500. The area under the curve (AUC) values ​​for these markers were as follows: Tau, 0.938 (95% CI 0.877-0.998); 4R Tau, 0.765 (95% CI 0.636-0.893); pTau181, 0.800 (95% CI 0.681-0.919); pTau231, 0.644 (95% CI 0.495-0.792); pTau396, 0.634 (95% CI 0.481-0.787) ( Figure 2 Middle K).

[0062] In differentiating PSP from PD, the AUC values ​​of these neuronal EVs were Tau, 0.912 (95% CI 0.841-0.983); 4R Tau, 0.795 (95% CI 0.678-0.912); pTau181, 0.730 (95% CI 0.593-0.867); pTau231, 0.603 (95% CI 0.454-0.752); pTau396, 0.519 (95% CI 0.366-0.673) ( Figure 2 The cut-off values ​​(unit: cells / mL) are as follows: Tau, 1,039,500; 4R Tau, 1,152,000; pTau181, 2,584,500.

[0063] 3. Results of multivariate logistic regression model analysis

[0064] (1) Differentiate between PSP and HC

[0065] The equation variables of the multivariate logistic regression model established in the present invention for distinguishing PSP from HC are shown in Table 1, and the risk value calculation equation is as follows:

[0066] Risk value D1 = 0.0000054431 × C tau +0.0000043461×C 4RTau +0.0000004477×C pTau181 +0.0000032264×C pTau231 -0.0000001555×C pTau396 ;

[0067] In the formula, C tau , C 4R Tau , C pTau181 , C pTau231 and C pTau396 Represents the concentration of neuronal-derived EVs containing Tau, 4R Tau, pTau181, pTau231, or pTau396 in plasma, respectively.

[0068] Table 1 Equation variables of the multivariate logistic regression model for distinguishing PSP from HC

[0069] Markers B Standard error Wald Degrees of Freedom Significance Exp(B) Tau 0.0000054431 0.0000020812 6.840 1 0.0089118534 1.0000054431 4Rtau 0.0000043461 0.0000028399 2.342 1 0.1259263804 1.0000043461 pTau181 0.0000004477 0.0000004736 0.894 1 0.3445216510 1.0000004477 pTau231 0.0000032264 0.0000028035 1.324 1 0.2497914587 1.0000032264 pTau396 -0.0000001555 0.0000003090 0.253 1 0.6149223604 0.9999998445 constant -14.9399425709 6.0845472380 6.029 1 0.0140731422 0.0000003248

[0070] The multivariate logistic regression model incorporating these neuronal EVs carrying Tau had an AUC of 0.974 (95% CI 0.941-1.000) and a cut-off value (unit: EVs / mL) of 0.6267, indicating a sensitivity of 95.7% and a specificity of 90.0% ( Figure 2 Middle L).

[0071] (2) Differentiate between PSP and PD

[0072] The equation variables of the multivariate logistic regression model established in the present invention for distinguishing PSP from PD are shown in Table 2, and the risk value calculation equation is as follows:

[0073] Risk value D2 = 0.0000070751 × C tau +0.0000028567×C 4RTau +0.0000006862×C pTau181 +0.0000013800×C pTau231 -0.0000004485×CpTau396 ;

[0074] In the formula, C tau , C 4R Tau , C pTau181 , C pTau231 and C pTau396 Represents the concentration of neuronal-derived EVs containing Tau, 4R Tau, pTau181, pTau231, or pTau396 in plasma, respectively.

[0075] Table 2 Equation variables of the multivariate logistic regression model for distinguishing PSP from PD

[0076]

[0077]

[0078] The corresponding multivariate logistic regression model integrating these biomarkers had an AUC of 0.968 (95% CI 0.929-1.000), a cut-off value (unit: cells / mL) of 0.6398, a sensitivity of 96.3% and a specificity of 90.0% ( Figure 2 (in N).

[0079] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A biomarker for assisting the diagnosis of progressive supranuclear palsy, characterized in that: The biomarker is a neuron-derived extracellular vesicle carrying Tau deformed protein; The Tau deformed protein is 4R Tau or pTau181.

2. The biomarker according to claim 1, characterized in that A higher level of the biomarker in plasma indicates a higher risk of the patient suffering from progressive supranuclear palsy.

3. A biomarker for distinguishing progressive supranuclear palsy from Parkinson's disease, characterized in that: The biomarker is a neuron-derived extracellular vesicle carrying Tau deformed protein; The Tau deformed protein is 4R Tau or pTau181.

4. The biomarker according to claim 3, characterized in that A higher level of the biomarker in plasma indicates that the patient has a higher risk of developing progressive supranuclear palsy among progressive supranuclear palsy and Parkinson's disease.

5. A diagnostic model for progressive supranuclear palsy ND Tau / Tau, characterized in that: The diagnostic model ND Tau / Tau uses the plasma concentration of neuron-derived extracellular vesicles carrying Tau protein and the plasma concentration of total extracellular vesicles carrying Tau protein as input variables; The ratio diagnostic model is: risk value R = plasma concentration of neuron-derived extracellular vesicles carrying Tau protein / plasma concentration of total extracellular vesicles carrying Tau protein; A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy.

6. A diagnostic model for progressive supranuclear palsy ND pTau231 / pTau231, characterized in that: The diagnostic model ND pTau231 / pTau231 uses the plasma concentration of neuron-derived extracellular vesicles carrying pTau231 protein and the plasma concentration of total extracellular vesicles carrying pTau231 protein as input variables; The ratio diagnostic model is risk value R=plasma concentration of neuron-derived extracellular vesicles carrying pTau231 protein / plasma concentration of total extracellular vesicles carrying pTau231 protein; A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy.

7. A diagnostic model ND 4R Tau / 4R Tau for differentiating progressive supranuclear palsy from Parkinson's disease, characterized in that: The diagnostic model ND 4R Tau / 4R Tau uses the plasma concentration of neuron-derived extracellular vesicles carrying 4R Tau protein and the plasma concentration of total extracellular vesicles carrying 4RTau protein as input variables; The ratio diagnostic model is: risk value R = plasma concentration of neuron-derived extracellular vesicles carrying 4R Tau protein / plasma concentration of total extracellular vesicles carrying 4R Tau protein; A higher value of the risk value R indicates that the patient has a higher risk of suffering from progressive supranuclear palsy among progressive supranuclear palsy and Parkinson's disease.

8. A multivariate logistic regression model for assisting the diagnosis of progressive supranuclear palsy, characterized in that: When used to distinguish patients with progressive supranuclear palsy from normal subjects, the risk value of the multivariate logistic regression model was D1 = 0.0000054431 × C tau +0.0000043461×C 4RTau +0.0000004477×C pTau181 +0.0000032264×C pTau231 -0.0000001555×C pTau396 ; When used to distinguish patients with progressive supranuclear palsy from patients with Parkinson's disease, the risk value of the multivariate logistic regression model was D2 = 0.0000070751 × C tau +0.0000028567×C 4RTau +0.0000006862×C pTau181 +0.0000013800×C pTau231 -0.0000004485×C pTau396 ; The C tau , said C 4RTau , said C pTau181 , said C pTau231 and the C pTau396 Represents the concentration of neuronal-derived extracellular vesicles containing Tau, 4RTau, pTau181, pTau231 or pTau396 in plasma, respectively.

9. Use of a reagent for detecting the content of the biomarker according to claim 1 in plasma in the preparation of a kit for auxiliary diagnosis of progressive supranuclear palsy.

10. The use according to claim 9, characterized in that: The reagent is a reagent for nanoscale flow cytometry detection.

Images