Plasma lipid markers for detecting elevated cerebrospinal fluid leukocytes in vkh and uses thereof

Abstract

The application discloses a plasma lipid marker for detecting a cerebrospinal fluid leukocyte increase in VKH and application thereof, and particularly comprises any one or more of the following: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4). The application fills the blank of a marker for non-invasive prediction of cerebrospinal fluid cell abnormalities in VKH syndrome, and can provide a new objective molecular index for realizing early non-invasive auxiliary diagnosis of the disease in clinic.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to plasma lipid markers for detecting elevated white blood cell counts in cerebrospinal fluid in patients with VKH and their applications. Background Technology

[0002] Vogt-Koyanagi-Harada syndrome (VKH) is an autoimmune disease characterized by bilateral granulomatous uveitis that can affect multiple systems, including the eye, meninges, inner ear, and skin. VKH can be divided into two key stages: the newly diagnosed acute phase (early stage) and the chronic relapsing phase (late stage). If early diagnosis of VKH is delayed or initial treatment is inadequate, the disease is highly likely to progress to the chronic relapsing phase. Once in this stage, treatment becomes significantly more difficult, the incidence of complications rises dramatically, and it often leads to irreversible and severe visual impairment. [1] In recent years, increasing evidence suggests that there is a clear window of opportunity for treatment in early-stage VKH, and timely intervention at this stage may even achieve clinical cure, highlighting the important clinical value of early accurate diagnosis and immediate initiation of standardized treatment. [2-4] .

[0003] However, early ocular manifestations of VKH are often nonspecific, or even asymptomatic, making it easily confused with posterior scleritis, acute posterior multifocal squamous pigment epithelial lesions, hypertensive retinopathy, and multifocal central serous chorioretinopathy, leading to misdiagnosis and missed diagnosis. Elevated leukocytes in cerebrospinal fluid are a relatively specific early sign of VKH, which can aid in early diagnosis; however, lumbar puncture is an invasive procedure with inherent risks of infection and nerve damage, significantly limiting its clinical application. A retrospective study of 76 patients diagnosed with VKH showed an initial misdiagnosis rate as high as 9%, further highlighting significant deficiencies in the current clinical diagnostic system. [5] Therefore, establishing accurate, objective, and non-invasive novel diagnostic biomarkers to assist in the early identification and precise diagnosis of VKH has become a key issue that urgently needs to be addressed in clinical practice.

[0004] Lipid metabolism plays a crucial regulatory role in autoimmune and inflammation-related diseases. Peripheral blood lipid profiles, with their readily available and repeatable characteristics, hold promise as ideal biomarkers reflecting disease pathology. This invention utilizes lipidomics analysis of plasma from newly diagnosed acute (early) VKH patients and healthy controls to screen for nine lipid substances significantly associated with cerebrospinal fluid leukocytosis. Both individual and combined detection of these lipids demonstrated good diagnostic efficacy, providing a non-invasive, early, and objective diagnostic method for VKH. This effectively compensates for the shortcomings of existing diagnostic methods and has significant clinical application value. Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide a plasma lipid biomarker for detecting elevated cerebrospinal fluid white blood cell count in VKH and its application. This invention aims to solve the following technical problems: First, early clinical manifestations of VKH are atypical and lack specific external indicators, making it easy to miss or misdiagnose clinically, and difficult to clarify the condition in the early stages; Second, elevated cerebrospinal fluid white blood cell count is a key characteristic indicator of this disease, but it requires invasive lumbar puncture for sampling, which carries high operational risks, poor patient compliance, and cannot be quickly implemented or repeatedly monitored; Third, there is a lack of peripheral blood biomarkers that can non-invasively and indirectly reflect elevated cerebrospinal fluid white blood cell counts, making it impossible to achieve non-invasive prediction and rapid screening, and failing to meet the actual needs of early clinical auxiliary diagnosis.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] 1. Plasma lipid markers for detecting elevated leukocytes in cerebrospinal fluid during VKH, including one or more of the following: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4).

[0008] Preferred combinations are those of the following nine lipids: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4).

[0009] Preferably, the plasma lipid marker content in the plasma of VKH patients is significantly negatively correlated with the cerebrospinal fluid white blood cell count.

[0010] 2. Application of the aforementioned plasma lipid markers in the preparation of a cerebrospinal fluid leukocyte detection kit.

[0011] Preferably, the test sample for the kit is peripheral blood.

[0012] Preferably, the kit is used for early screening or diagnosis of VKH syndrome.

[0013] Preferably, the kit is a non-invasive screening or diagnostic kit.

[0014] Preferably, the kit is able to distinguish between patients with VKH syndrome and healthy individuals.

[0015] 3. Application of the aforementioned plasma lipid markers in the preparation of early screening or diagnostic kits for VKH syndrome.

[0016] 4. Kit, containing reagents for detecting the levels of the aforementioned plasma lipid markers.

[0017] Preferably, the kit includes lipid extraction reagents and chromatographic mobile phase reagents; the lipid extraction reagents include: methanol, methyl tert-butyl ether solution containing internal standard, and dichloromethane / methanol mixture (volume ratio 1:1); the chromatographic mobile phase reagents include: mobile phase A for reversed-phase separation: a water / acetonitrile / methanol mixture containing 7 mmol / L ammonium acetate (volume ratio 1:1:1), and mobile phase B: an isopropanol solution containing 7 mmol / L ammonium acetate; mobile phase A for HILIC separation: a water / acetonitrile mixture containing 2 mmol / L ammonium acetate (volume ratio 1:1), and mobile phase B: a dichloromethane / acetonitrile mixture containing 2 mmol / L ammonium acetate (volume ratio 1:13).

[0018] More preferably, the method of using the kit is as follows: Take 50 μL of plasma, add 200 μL of methanol and mix well, add 750 μL of methyl tert-butyl ether solution containing internal standard, add pure water to separate the phase, collect the organic phase and dry it, reconstitute it with dichloromethane / methanol mixture, and then perform MRM mode detection by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] This invention provides plasma lipid biomarkers for detecting elevated cerebrospinal fluid leukocytes in VKH syndrome and their applications. Specifically, these biomarkers include one or more of the following: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (0-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4). This invention fills the gap in biomarkers for non-invasive prediction of cerebrospinal fluid cellular abnormalities in VKH syndrome and can provide a novel objective molecular indicator for early non-invasive auxiliary diagnosis of this disease in clinical practice.

[0021] This invention utilizes ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS / MS) to perform lipidomics analysis on the plasma of patients with early-stage VKH and healthy individuals, obtaining lipid metabolite information. Based on this, the invention employs fold difference screening, orthogonal partial least squares discriminant analysis (OPLS-DA), and correlation analysis to screen nine lipid metabolites as potential lipid metabolites for VKH diagnosis. The diagnostic efficacy of these lipid biomarkers is verified by calculating the receiver operating characteristic (ROC) curve and area under the curve (AUC).

[0022] The above combination of lipid biomarkers exhibits excellent predictive efficacy, with good sensitivity and specificity. It can accurately distinguish between subjects with normal and elevated cerebrospinal fluid white blood cell counts, assisting clinicians in quickly assessing the condition, reducing the probability of missed or misdiagnosed cases, and providing a reliable basis for clinical diagnosis and treatment decisions.

[0023] This invention uses peripheral blood plasma as the test sample, which is simple to obtain, minimally invasive and painless, avoiding the drawbacks of lumbar puncture such as invasive damage and complication risks. It has high patient acceptance and is suitable for clinical batch testing, early screening and dynamic follow-up of the condition. Attached Figure Description

[0024] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the following figures are provided for illustration:

[0025] Figure 1 This is a lipid volcano plot showing the differences between VKH patients and healthy individuals;

[0026] Figure 2 This is a correlation analysis between plasma lipid metabolite levels and cerebrospinal fluid leukocytes in VKH patients;

[0027] Figure 3 It is the ROC curve of lipid metabolites;

[0028] Figure 4 It is a combined diagnostic ROC curve of 9 lipid metabolites. Detailed Implementation

[0029] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

[0030] Variable Importance in Projection (VIP): Primarily used in multivariate statistical models such as OPLS-DA, this quantifies the contribution of each independent variable to the construction of the model's projection space and the explanation of the dependent variable's variation. In metabolomics OPLS-DA models, a VIP > 1 threshold is typically used to screen differentially expressed lipids and metabolites that significantly contribute to sample grouping.

[0031] False Discovery Rate (FDR): A statistical correction metric for multiple hypothesis testing scenarios, referring to the proportion of false positive results (incorrect rejection of the null hypothesis) among all test results judged as significantly positive.

[0032] P-value: The core statistic for hypothesis testing, which indicates the probability of obtaining the current observed data or more extreme data given that the null hypothesis is true.

[0033] MetaboAnalyst: A free, web-based metabolomics data analysis and annotation platform.

[0034] Example 1:

[0035] 1. Collection of plasma samples

[0036] This study was conducted in strict accordance with the Declaration of Helsinki and approved by the Ethics Committee of the First Affiliated Hospital of Chongqing Medical University. From February 2022 to December 2025, the study recruited 26 patients with newly diagnosed acute (early) VKH syndrome and 39 healthy controls at the First Affiliated Hospital of Chongqing Medical University. All participants were voluntary, and all signed written informed consent forms after fully understanding the study content, risks, and rights. Prior to blood collection, none of the patients had received systemic glucocorticoids, immunosuppressants, or biologics. Patient diagnosis strictly followed the VKH diagnostic criteria revised by the International Committee, combined with the clinical diagnostic criteria established by our research group. All patients presented with early ocular manifestations, including diffuse choroiditis and exudative retinal detachment, accompanied by neurological symptoms and signs. Exclusion criteria included hypertension, cardiovascular disease, diabetes, hepatitis, a history of tuberculosis, or other autoimmune diseases. There were no significant differences in age, sex, or body mass index among all participants.

[0037] 2. Plasma sample pretreatment

[0038] Blood samples were collected in coagulation tubes and allowed to stand at room temperature for 30 minutes before being centrifuged at 3000 rpm for 10 minutes to separate the plasma. The separated plasma was stored at -80°C, avoiding repeated freeze-thaw cycles throughout the process.

[0039] Lipidomics analysis was performed using the methyl tert-butyl ether (MTBE) method: 50 μL of plasma was added to 200 μL of methanol and mixed well, followed by 750 μL of MTBE solution containing internal standard. After shaking and mixing, an appropriate amount of pure water was added to induce phase separation. After centrifugation, the upper organic phase was collected. The organic phase was then evaporated and dried in a vacuum concentrator, and subsequently reconstituted with a dichloromethane / methanol mixed solution at a volume ratio of 1:1 (v / v). The reconstituted phase was then used for subsequent quantitative lipidomics analysis.

[0040] 3. Quantitative lipid analysis

[0041] Quantitative lipid analysis was performed using an Agilent 1290 Infinity II ultra-high performance liquid chromatography system coupled with a SCIEXTripleQuad™ 7500+ triple quadrupole mass spectrometer.

[0042] Reversed-phase separation was performed using a Kinetex C18 column (2.1 × 100 mm, 2.6 μm particle size). Mobile phase A was a water / acetonitrile / methanol mixture containing 7 mmol / L ammonium acetate at a volume ratio of 1:1:1 (v / v / v), and mobile phase B was an isopropanol solution containing 7 mmol / L ammonium acetate. Hydrophilic interaction chromatography (HILIC) separation was performed using a Luna NH2 column (2.0 × 100 mm, 3 μm particle size). Mobile phase A was a water / acetonitrile mixture containing 2 mmol / L ammonium acetate at a volume ratio of 1:1 (v / v), and mobile phase B was a dichloromethane / acetonitrile mixture containing 2 mmol / L ammonium acetate at a volume ratio of 1:13 (v / v).

[0043] Mass spectrometry was performed in multiple reaction monitoring (MRM) mode with an ion spray voltage of ±3000 V, a curtain gas pressure of 35 psi, and ion source gas pressures of 40 psi and 70 psi (high-purity nitrogen) for ion source gas 1 and gas 2 (high-purity nitrogen), respectively. The ion source temperature was 325 °C in positive mode and 550 °C in negative mode. Data acquisition and quantitative analysis were performed using SCIEXOS software (version 3.3.1) and BIOTREEBioBud software (version 2.0.4). After acquisition, the raw liquid chromatography-tandem mass spectrometry data were preprocessed. Lipid peaks with a missing rate exceeding 50% in any given set were removed, and the remaining missing values ​​were filled with half the minimum value of the corresponding peak, retaining a total of 1186 lipid peaks for subsequent data analysis.

[0044] 4. Data Analysis

[0045] Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis were performed using SIMCA version 18.0.1 software. PCA was used as an unsupervised analysis method for visualizing the overall distribution of the sample. Figure 1A total of 346 significantly differentially expressed lipids (VIP > 1, P < 0.05) were identified. OPLS-DA was used to evaluate the separation effect between groups and to screen differentially expressed lipid metabolites. The R² and Q² values ​​obtained by 7-fold cross-validation were used to evaluate the fit of the OPLS-DA model, and the robustness of the model was verified by 1000 permutations to eliminate the risk of overfitting. The contribution of each variable was evaluated based on the variable projection importance (VIP) score of the OPLS-DA model. Lipid metabolites with VIP > 1 and P < 0.05 (FDR corrected) were defined as significantly differentially expressed molecules, and 262 differentially expressed lipids were finally screened. MetaboAnalyst software was used for metabolic pathway enrichment analysis to identify the key biological pathways associated with differentially expressed metabolites and lipids.

[0046] After screening out 262 lipids, correlation analysis between lipids and cerebrospinal fluid white blood cell count was performed. Spearman's test was used to retain 9 lipids with P < 0.05. The specific P values ​​and r values ​​are shown in Table 1.

[0047] Table 1

[0048] Phosphatidylethanolamine 16:0 / 22:4 0.876 0.764--0.988 -0.5929 0.002869 [-0.8077, -0.2391] Alkyl ether phosphatidylethanolamine O-18:0 / 22:4 0.720 0.586--0.854 -0.5375 0.00816 [-0.7775, -0.161] alkenyl ether phosphatidylethanolamine P-20:0 / 22:4 0.833 0.721--0.945 -0.5277 0.009662 [-0.772, -0.1476] Phosphatidylethanolamine 19:0 / 22:4 0.707 0.569--0.845 -0.5198 0.011021 [-0.7675, -0.1369] alkenyl ether phosphatidylethanolamine P-18:0 / 22:4 0.822 0.713--0.931 -0.501 0.014887 [-0.7569, -0.1119] Phosphatidylethanolamine 20:3 / 22:4 0.776 0.651--0.902 -0.4901 0.017589 [-0.7507, -0.0976] Phosphatidylcholine 14:0 / 14:0 0.726 0.592--0.860 -0.4881 0.018121 [-0.7495, -0.095] alkenyl ether phosphatidylethanolamine P-18:0 / 22:5 0.791 0.672--0.910 -0.4358 0.037661 [-0.7189, -0.0288] alkenyl ether phosphatidylethanolamine P-16:0 / 22:4 0.855 0.745--0.965 -0.4229 0.044362 [-0.7111, -0.013]

[0049] 5. Correlation analysis

[0050] Depend on Figure 2 Nine glycerophospholipids showed a significant negative correlation with cerebrospinal fluid leukocyte count, including phosphatidylethanolamine (16:0 / 22:4), alkyl ether phosphatidylethanolamine (O-18:0 / 22:4), alkenyl ether phosphatidylethanolamine (P-20:0 / 22:4), phosphatidylethanolamine (19:0 / 22:4), alkenyl ether phosphatidylethanolamine (P-18:0 / 22:4), phosphatidylethanolamine (20:3 / 22:4), phosphatidylcholine (14:0 / 14:0), alkenyl ether phosphatidylethanolamine (P-18:0 / 22:5), and alkenyl ether phosphatidylethanolamine (P-16:0 / 22:4).

[0051] Depend on Figure 3 As can be seen, the diagnostic potential of the above nine lipids was evaluated by ROC curve analysis, and their AUC ranged from 0.71 to 0.88, indicating that they have good distinguishing power.

[0052] Depend on Figure 4 The AUC for the combined diagnosis of the nine lipids was 0.889. Both the individual and combined detection of these lipids demonstrated good diagnostic efficacy.

[0053] Example 2:

[0054] Diagnostic kit for assessing central nervous system involvement in early VKH patients

[0055] A diagnostic kit for assessing central nervous system involvement in early VKH patients, consisting of one or more of the following: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4).

[0056] Based on the above nine lipid metabolite combinations, a VKH early non-invasive diagnostic kit was prepared. The method of use is as follows: take a plasma sample from the subject to be tested, extract and detect lipids according to the instructions, measure the expression level of the target lipid metabolite, compare it with the preset threshold, and determine whether it is VKH positive.

[0057] Specifically, the kit includes lipid extraction reagents and chromatographic mobile phase reagents; the lipid extraction reagents include: methanol, methyl tert-butyl ether solution containing internal standard, and dichloromethane / methanol mixture (volume ratio 1:1); the chromatographic mobile phase reagents include: mobile phase A for reversed-phase separation: a water / acetonitrile / methanol mixture containing 7 mmol / L ammonium acetate (volume ratio 1:1:1), and mobile phase B: an isopropanol solution containing 7 mmol / L ammonium acetate; mobile phase A for HILIC separation: a water / acetonitrile mixture containing 2 mmol / L ammonium acetate (volume ratio 1:1), and mobile phase B: a dichloromethane / acetonitrile mixture containing 2 mmol / L ammonium acetate (volume ratio 1:13).

[0058] The method of using this kit is as follows: Take 50 μL of plasma, add 200 μL of methanol and mix well, add 750 μL of methyl tert-butyl ether solution containing internal standard, add pure water to separate the phase, collect the organic phase and dry it, reconstitute it with dichloromethane / methanol mixture, and then perform MRM mode detection by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.

[0059] The specific methods for lipid extraction are as follows:

[0060] 2-1) Remove the centrifuge tube containing plasma from the -80℃ ultra-low temperature freezer, place it on ice to thaw slowly, and gently vortex to mix after thawing, avoiding violent shaking that could lead to lipid degradation.

[0061] 2-2) Use a pipette to draw 50 μL of thawed plasma and add it to a new 1.5 mL sterile centrifuge tube. Add 200 μL of methanol, place the centrifuge tube on a vortex mixer, and shake for 30 seconds to mix thoroughly and achieve plasma protein membrane rupture.

[0062] 2-3) Add 750 μL of MTBE (methyl tert-butyl ether) solution containing internal standard to the centrifuge tube, vortex for 1 minute to fully dissolve the lipids in the MTBE organic phase.

[0063] 2-4) Add an appropriate amount of ultrapure water to the centrifuge tube, vortex for 30 seconds to mix, and let stand at room temperature for 5 minutes to induce the separation of the aqueous and organic phases. Then place the centrifuge tube in a centrifuge, adjust the speed to a suitable speed (usually 12000 rpm), and centrifuge for 5 minutes to completely separate the two phases.

[0064] 2-5) After centrifugation, slowly aspirate the upper transparent organic phase (the lipid phase) with a pipette and transfer it to a new sterile 1.5mL centrifuge tube, avoiding aspirating the lower aqueous phase and the intermediate impurity layer.

[0065] 2-6) Place the centrifuge tube containing the organic phase into a vacuum concentrator, set appropriate conditions (temperature ≤30℃, vacuum degree according to instrument calibration), evaporate and dry until the organic phase is completely volatilized to obtain lipid powder.

[0066] 2-7) Add an appropriate amount of dichloromethane / methanol mixed solution (volume ratio 1:1, v / v) to the dried centrifuge tube, vortex for 1 minute to completely reconstitute the lipid powder. The reconstituted solution is the lipid extract, which is used for subsequent quantitative lipidomics analysis.

[0067] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made to it in form and detail without departing from the scope defined by the claims of the present invention.

[0068] References

[0069] [1] Urzua CA, Herbort C, Jr., Valenzuela RA, et al. Initial-onsetacute and chronic recurrent stages are two distinctive courses of Vogt-Koyanagi-Harada disease. J Ophthalmic Inflamm Infect 2020; 10: 23. 20200914.DOI: 10.1186 / s12348-020-00214-2.

[0070] [2] Herbort CP, Jr., Abu El Asrar AM, Takeuchi M, et al. Catching thetherapeutic window of opportunity in early initial-onset Vogt-Koyanagi-Haradauveitis can cure the disease. Int Ophthalmol 2019; 39: 1419-1425. 20180611.DOI: 10.1007 / s10792-018-0949-4.

[0071] [3] Papasavvas I, Tugal-Tutkun I and Herbort CP, Jr. Vogt-Koyanagi-Harada is a Curable Autoimmune Disease: Early Diagnosis and Immediate DualSteroidal and Non-Steroidal Immunosuppression are Crucial Prerequisites. JCurr Ophthalmol 2020; 32: 310-314. 20201212. DOI: 10.4103 / joco.Joco_190_20.

[0072] [4] Yang P, Ye Z, Du L, et al. Novel treatment regimen of Vogt-Koyanagi-Harada disease with a reduced dose of corticosteroids combined withimmunosuppressive agents. Curr Eye Res 2018; 43: 254-261. 20171107. DOI:10.1080 / 02713683.2017.1383444.

[0073] [5] Shoughy SS and Tabbara KF. Initial misdiagnosis of Vogt-Koyanagi-Harada disease. Saudi J Ophthalmol 2019; 33: 52-55. 20181207. DOI: 10.1016 / j.sjopt.2018.11.006.

Claims

1. Plasma lipid markers for detecting elevated cerebrospinal fluid leukocytes in VKH, characterized in that, Including any one or more of the following: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4).

2. The plasma lipid marker according to claim 1, characterized in that, It is a combination of the following 9 lipids: phosphatidylethanolamine PE (16:0 / 22:4), phosphatidylethanolamine PE (O-18:0 / 22:4), phosphatidylethanolamine PE (P-20:0 / 22:4), phosphatidylethanolamine PE (19:0 / 22:4), phosphatidylethanolamine PE (P-18:0 / 22:4), phosphatidylethanolamine PE (20:3 / 22:4), phosphatidylcholine PC (14:0 / 14:0), phosphatidylethanolamine PE (P-18:0 / 22:5), and phosphatidylethanolamine PE (P-16:0 / 22:4).

3. The plasma lipid marker according to claim 1, characterized in that, The levels of the plasma lipid markers in the plasma of VKH patients showed a significant negative correlation with the cerebrospinal fluid white blood cell count.

4. The use of the plasma lipid markers of claim 1 in the preparation of a cerebrospinal fluid leukocyte detection kit.

5. The application according to claim 4, characterized in that, The test sample for this kit is peripheral blood.

6. The application according to claim 4, characterized in that, The kit is used for early screening or diagnosis of VKH syndrome.

7. The application according to claim 4, characterized in that, The kit is a non-invasive screening or diagnostic kit.

8. The application according to claim 4, characterized in that, The kit can distinguish between patients with VKH syndrome and healthy individuals.

9. The use of the plasma lipid biomarker of claim 1 in the preparation of a kit for early screening or diagnosis of VKH syndrome.

10. A reagent kit, characterized in that, It includes reagents for detecting the content of the plasma lipid markers of claim 1.

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