Combined lipid biomarkers for aiding diagnosis of schizophrenia and use thereof

Abstract

The application belongs to the technical field of biotechnology, and particularly relates to a combined lipid biomarker for auxiliary diagnosis of schizophrenia and application thereof. The application realizes auxiliary diagnosis of schizophrenia by detecting content changes of specific molecular subtypes TG(40:0-FA16:0), PE(18:1 / 18:3), PE(18:2 / 18:2) and / or PE(18:1 / 18:2) of triglyceride and phosphatidylethanolamine in peripheral blood, so as to overcome the defects that the existing diagnosis of schizophrenia mainly depends on clinical symptom evaluation, and is subjective and lacks consistency.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and specifically relates to peripheral blood biomarkers for the objective diagnosis of schizophrenia. Background Technology

[0002] Schizophrenia (SCZ) is a chronic, highly relapsing, and highly disabling severe mental disorder. Its core clinical features include positive symptoms, negative symptoms, and varying degrees of cognitive impairment. The disease typically begins in young adulthood, and its main clinical manifestations include mood disturbances, abnormal thinking and behavior, reduced volition, and cognitive impairment. Schizophrenia has a long-term and profound impact on patients' social functioning and quality of life, and its tendency to relapse places a heavy economic burden on patients' families and puts significant pressure on public safety and medical resources. Epidemiological studies indicate that the lifetime prevalence of schizophrenia in the global population is approximately 1%, and its overall disease burden continues to rise due to demographic changes and increasing mental health needs. Currently, the clinical diagnosis of schizophrenia mainly relies on symptomatological interviews and scale measurements by psychiatrists, lacking objective, quantifiable, and easily generalized biological indicators. Because early clinical symptoms are often atypical and exhibit significant individual differences, some patients are only diagnosed in the middle to late stages of the disease, thus missing the optimal intervention window.

[0003] Although the exact pathogenesis of schizophrenia is not fully understood, a growing body of research indicates that abnormal lipid metabolism plays a crucial role in its development and progression. Lipids are not only an important form of energy storage and supply for the body, but also key components of cell membrane structure and various signal transduction processes. Disorders of lipid metabolism can disrupt the structural and functional integrity of the blood-brain barrier through pathways such as inducing chronic inflammatory responses, enhanced oxidative stress, and mitochondrial dysfunction, thereby affecting central nervous system homeostasis and participating in the pathophysiological processes associated with schizophrenia.

[0004] Therefore, systematic analysis of specific lipid molecule types and their combination characteristics, and the development of stable, reproducible, and easily obtainable peripheral biomarkers, are of great significance for early screening, auxiliary diagnosis, and disease risk assessment of schizophrenia. Summary of the Invention

[0005] The purpose of this invention is to provide a peripheral blood lipid biomarker for the objective diagnosis of schizophrenia. By detecting changes in the levels of specific molecular subtypes of triglycerides and phosphatidylethanolamine in peripheral blood, triglycerides (40:0-FA16:0) (TG(40:0-FA16:0)), phosphatidylethanolamine (18:1 / 18:3) (PE(18:1 / 18:3)), phosphatidylethanolamine (18:2 / 18:2) (PE(18:2 / 18:2)) and / or phosphatidylethanolamine (18:1 / 18:2) (PE(18:1 / 18:2)), the invention aims to assist in the diagnosis of schizophrenia, thereby overcoming the shortcomings of existing schizophrenia diagnoses, which mainly rely on clinical symptom assessment, are highly subjective, and lack consistency.

[0006] TG(40:0-FA16:0) refers to a specific subtype of triglyceride molecule with a total of 40 carbon atoms, 0 double bonds, and one fatty acid chain being 16:0; PE(18:1 / 18:3) refers to a monounsaturated fatty acid with 18 carbon atoms and one carbon-carbon double bond attached to the sn-1 position of the PE glycerol backbone, and a polyunsaturated fatty acid with 18 carbon atoms and three carbon-carbon double bonds attached to the sn-2 position; PE(18:2 / 18:2) refers to a fatty acid with 18 carbon atoms and two carbon-carbon double bonds attached to both the sn-1 and sn-2 positions of the PE glycerol backbone; PE(18:1 / 18:2) refers to a PE glycerol backbone with an 18:1 fatty acid attached to the sn-1 position and an 18:2 fatty acid attached to the sn-2 position.

[0007] On one hand, this invention provides the application of any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) as biomarkers in the preparation of diagnostic products for schizophrenia. This invention has discovered significant differences in the above-mentioned lipid molecules in the peripheral blood of schizophrenia patients and normal individuals. These differences are sufficient to accurately distinguish between schizophrenia patients and normal individuals, making them suitable as biomarkers for detecting schizophrenia-related conditions. These biomarkers exhibit high accuracy, high sensitivity, and strong specificity. This invention also compares the diagnostic efficacy of single biomarkers with combinations of four biomarkers. The results demonstrate that combined use significantly improves diagnostic performance (AUC reaches 0.909), superior to any single indicator; therefore, combined application has a prominent synergistic effect.

[0008] This invention has determined that the levels of TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2), and PE (18:1 / 18:2) in human peripheral blood are significantly associated with schizophrenia. Therefore, the risk of schizophrenia can be assessed by detecting the levels of one or more of these lipid molecules in peripheral blood. As for the specific detection method, it can be any method disclosed in the prior art, such as chromatography, spectroscopy, mass spectrometry, liquid chromatography-mass spectrometry, or chemical analysis methods. This invention uses liquid chromatography-mass spectrometry for detection, but it is not limited to this method.

[0009] On the other hand, the present invention also provides the use of reagents for detecting any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the preparation of diagnostic products for schizophrenia.

[0010] In some embodiments, the product includes reagents for detecting any one or more of the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0011] In some embodiments, the product includes a reagent for detecting any one of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0012] In some embodiments, the product includes reagents for detecting triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0013] In some embodiments, the triglyceride (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), or phosphatidylethanolamine (18:1 / 18:2) samples are derived from peripheral blood.

[0014] In some embodiments, the reagents used to detect the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and / or phosphatidylethanolamine (18:1 / 18:2) may be the same or different.

[0015] In some implementations, the reagent is a reagent used for detection by chromatographic, spectroscopic, mass spectrometric, liquid chromatography-mass spectrometry, or chemical analysis methods.

[0016] In some embodiments, the product includes a C18 column, mobile phase A being 60 / 40 acetonitrile / water containing 5 mM ammonium acetate, and mobile phase B being 90 / 10 isopropanol / acetonitrile containing 5 mM ammonium acetate.

[0017] In some implementations, the flow rate in the liquid chromatography-mass spectrometry detection is 0.3 mL / min, and the elution program is 0.0 min B phase 30%, 3.0 min B phase 70%, 12.5 min B phase 95%, 12.6 min B phase 30%, and stop operation at 14 min.

[0018] In some embodiments, the liquid chromatography-mass spectrometry (LC-MS) uses a Heated ESI electrospray ionization source with a positive ion ionization voltage of 3.5 kV, a negative ion ionization voltage of -3.0 kV, an ion source temperature of 350°C, a sheath gas flow rate of 40 Arb, an auxiliary gas flow rate of 15 Arb, and a collision gas pressure of 2.0 mTorr.

[0019] In some implementations, the product is a reagent kit.

[0020] In some implementation schemes, the diagnostic criteria are as follows: when the levels of any one or more of the following in the peripheral blood of the subject are significantly lower than those of healthy individuals, the subject is considered a suspected patient of schizophrenia.

[0021] In some implementation schemes, the judgment criteria are as follows: when the levels of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the peripheral blood of the subject are all significantly lower than those in healthy individuals, the subject is judged to be a suspected patient of schizophrenia.

[0022] In some implementations, "significantly lower than" means p < 0.0001.

[0023] In some implementations, triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) are used as biomarkers in combination.

[0024] In another aspect, the present invention also provides a diagnostic kit for schizophrenia, wherein the kit includes reagents for detecting any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0025] In some embodiments, the kit includes reagents for detecting any one of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0026] In some embodiments, the kit includes reagents for detecting triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2).

[0027] In some embodiments, the triglyceride (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), or phosphatidylethanolamine (18:1 / 18:2) samples are derived from peripheral blood.

[0028] In some embodiments, the reagents used to detect the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and / or phosphatidylethanolamine (18:1 / 18:2) may be the same or different.

[0029] In some implementations, the reagent is a reagent used for detection by chromatographic, spectroscopic, mass spectrometric, liquid chromatography-mass spectrometry, or chemical analysis methods.

[0030] In some embodiments, the kit includes a C18 column, mobile phase A being 60 / 40 acetonitrile / water containing 5 mM ammonium acetate, and mobile phase B being 90 / 10 isopropanol / acetonitrile containing 5 mM ammonium acetate.

[0031] In some implementations, the flow rate in the liquid chromatography-mass spectrometry detection is 0.3 mL / min, and the elution program is 0.0 min B phase 30%, 3.0 min B phase 70%, 12.5 min B phase 95%, 12.6 min B phase 30%, and stop operation at 14 min.

[0032] In some embodiments, the liquid chromatography-mass spectrometry (LC-MS) uses a Heated ESI electrospray ionization source with a positive ion ionization voltage of 3.5 kV, a negative ion ionization voltage of -3.0 kV, an ion source temperature of 350°C, a sheath gas flow rate of 40 Arb, an auxiliary gas flow rate of 15 Arb, and a collision gas pressure of 2.0 mTorr.

[0033] In some implementations, the diagnostic criteria for the kit are as follows: when the levels of any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the peripheral blood of the subject are significantly lower than those in healthy individuals, the subject is considered a suspected patient with schizophrenia.

[0034] In some implementation schemes, the judgment criteria are as follows: when the levels of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the peripheral blood of the subject are all significantly lower than those in healthy individuals, the subject is judged to be a suspected patient of schizophrenia.

[0035] In some implementations, "significantly lower than" means p < 0.0001.

[0036] Furthermore, the present invention also provides an auxiliary diagnostic method for schizophrenia, characterized by comprising the following steps: 1. Collect peripheral blood samples from the subjects and separate the plasma; 2. To detect the content of TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2) and / or PE (18:1 / 18:2) in the plasma; the preferred detection method is liquid chromatography-mass spectrometry. 3. Compare the test results with the reference values ​​of healthy controls. If the level of any one or more of the above lipid molecules in the peripheral blood of the test subject is significantly lower than that of healthy individuals (preferably p<0.0001), the subject is identified as a suspected patient with schizophrenia.

[0037] Advantages of this invention 1. This invention provides an objective biomarker system based on peripheral blood: Based on plasma-targeted lipidomics detection, this invention uses a combination of multiple biologically relevant lipid molecules for analysis, avoiding the high dependence of traditional schizophrenia diagnosis on symptom interviews and scale assessments, and improving the objectivity and reproducibility of diagnosis.

[0038] 2. Both single and combined indicators can significantly improve the diagnostic efficacy of schizophrenia: using a single indicator can achieve high diagnostic sensitivity and specificity, reduce detection costs and technical complexity, while the area under the ROC curve (AUC) of the combination of four lipid biomarkers as diagnostic indicators reaches 0.909 (95% CI: 0.850–0.961), with diagnostic sensitivity, specificity and accuracy of 95.6%, 82.2% and 87.5% respectively, showing comprehensive diagnostic performance superior to that of a single indicator.

[0039] 3. The detection method is mature and stable, with good clinical operability: The lipid molecules used in this invention can be detected by conventional mass spectrometry platforms such as liquid chromatography-mass spectrometry. The sample source is peripheral blood, and the collection method is simple and low-invasive, making it suitable for promotion and application in clinical and physical examination populations. Attached Figure Description

[0040] Figure 1 A scatter plot of lipid metabolites in peripheral blood of patients with schizophrenia and healthy controls is shown. The breakdown is as follows: TG: triglycerides; PE: phosphatidylethanolamine; PC: phosphatidylcholine; LysoPC: lysophosphatidylcholine; PI: phosphatidylinositol; FFA: free fatty acids; LysoPE: lysophosphatidylethanolamine; SM: sphingomyelin; HexCer: hexosylceramide; Cer: ceramide; CE: cholesterol esters; DG: diglycerides; LacCer: lactosylceramide; LysoPI: lysophosphatidylinositol; PG: phosphatidylglycerol; LysoPG: lysophosphatidylglycerol.

[0041] Figure 2 Volcano plots of lipid metabolites in peripheral blood of patients with schizophrenia and healthy controls are shown. In the figures, PE: phosphatidylethanolamine; FFA: free fatty acids; TG: triglycerides; PC: phosphatidylcholine; PI: phosphatidylinositol; PG: phosphatidylglycerol; CE: cholesterol esters; DG: diglycerides; Cer: ceramide; HexCer: hexosylceramide; LysoPE: lysophosphatidylethanolamine; LysoPC: lysophosphatidylcholine; LysoPG: lysophosphatidylglycerol; LysoPI: lysophosphatidylinositol; NotSig: not significant.

[0042] Figure 3 The levels of TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2), and PE (18:1 / 18:2) in peripheral blood of patients with schizophrenia and healthy controls are shown.

[0043] Figure 4The ROC curves of individual indicators and combinations of TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2), and PE (18:1 / 18:2) in peripheral blood of patients with schizophrenia and healthy controls are shown. Detailed Implementation

[0044] For the purpose of clarity and concise description, features are described herein as part of some identical or separate embodiments; however, it will be understood that the scope of the invention may include some embodiments having combinations of all or some of the described features. The technical solutions of the present invention will now be described clearly and completely. Obviously, based on the specific embodiments of the present invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of the present invention.

[0045] I. Definition Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0046] As used herein, the terms “contains,” “has,” “includes,” and “contains” should be interpreted as open-ended terms (i.e., meaning “including but not limited to”). “And / or” describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone. The character “ / ” generally indicates an “or” relationship between the preceding and following related objects, while the character “+” generally indicates an “and” relationship.

[0047] As used herein, the terms “optional,” “any,” “arbitrary,” or “any item” mean that the event or situation described below may, but does not have to, occur, including the circumstances in which the event or situation occurs or does not occur. As used herein, “a,” “an item,” and “a kind” are used to refer to one or more grammatical objects. “Multiple,” “various,” or “multiple items” as used herein refer to two or more grammatical objects, such as two, three, four, five, etc. The terms “one or more / a kind or more / an item or more” and “at least one / a kind / item” are used interchangeably.

[0048] As used herein, references to “some embodiments,” “some implementations,” or “some implementation schemes” describe a subset of all possible embodiments, but it is understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0049] As used herein, the term "about" indicates a range of ±20% of the following value. In some embodiments, the term "about" indicates a range of ±10% of the following value. In some embodiments, the term "about" indicates a range of ±5% of the following value.

[0050] II. Examples The present invention will be described in detail below through specific embodiments. It should be understood that the following embodiments are for explanation and illustration only and do not limit the scope of the present invention in any way.

[0051] Where specific methods, techniques, or conditions are not specified in the examples, they shall be performed in accordance with the methods, techniques, or conditions described in the literature in this field or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0052] Example 1: Sample Collection and Processing 1.2 Subject Screening Based on the approved ethical review protocol and diagnostic criteria, 86 healthy controls and 55 patients with schizophrenia were selected. The Positive and Negative Syndrome Scale (PANSS) was used to assess patient symptoms. Inclusion criteria referenced high-quality international cohort studies of first-episode psychosis, including the Early Intervention Services Development and Impact Assessment (EDEN) study in the UK, the Early Psychiatric Intervention Trial (OPUS) study in Denmark, the European Study for the Optimization of Schizophrenia Diagnosis and Management (OPTiMiSE), and the Early Psychiatric Prevention and Intervention Centre (EPPIC) in Australia. Screening was conducted using the Mini-International Neuropsychiatric Interview (MINI 7.0.2). Patients meeting the diagnostic criteria of schizophrenia, schizophrenia-like disorder, schizoaffective disorder, delusional disorder, and transient psychotic disorder as defined in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5), were: aged 13 to 50 years, right-handed, Han Chinese, with a junior high school education or higher, a total illness duration of less than 5 years, and a continuous antipsychotic medication use period of less than 2 weeks within the past year and a total previous antipsychotic medication use duration of less than 6 weeks.

[0053] Subjects in each group were excluded if they met any of the following criteria: current or past DSM-5 diagnostic criteria for substance abuse or substance dependence; intellectual disability or current serious physical illness; history of impaired consciousness lasting more than 5 minutes; contraindications to MRI; pregnant or lactating women; more than 3 ECT treatments in the past 12 months; or received transcranial direct current stimulation (tDCS), transcranial alternating current stimulation (tACS), or repetitive transcranial magnetic stimulation (rTMS) in the past 6 months.

[0054] Basic demographic information and symptom assessments for both groups are shown in Table 1. There were no statistically significant differences in age, sex, and body mass index (BMI) between patients with schizophrenia and healthy controls, indicating good baseline comparability.

[0055] Table 1. Basic demographic information of the subjects and assessment of patients' clinical symptoms Note: BMI, Body Mass Index; PANSS, Positive and Negative Syndrome Scale. χ2 The test is performed using the Mann-Whitney U test, and the rest are performed using the Mann-Whitney U test.

[0056] 1.2 Sample Collection Five milliliters of peripheral blood were collected from all subjects in EDTA anticoagulant tubes, centrifuged at 3000 rpm for 10 minutes, the supernatant plasma was separated, and immediately aliquoted and stored in an ultra-low temperature freezer at -80°C for later use.

[0057] Example 2: Lipid-targeted detection 2.1 Sample Preparation Accurately transfer 20 μL of blood sample to a 96-well plate, add 25 μL of isotope internal standard buffer (isotope internal standard concentration 2~1000 μg / mL), 215 μL of isopropanol (containing 10 mM ammonium acetate), vortex at 1500 rpm for 5 min, centrifuge at 5300 rpm for 20 min, and transfer the supernatant to a new 96-well plate for LC-MS / MS analysis.

[0058] 2.2 LC-MS / MS detection: Lipid targeting was performed using high performance liquid chromatography-tandem triple quadrupole mass spectrometry, and data were acquired using selected reaction monitoring (SRM) mode.

[0059] Liquid chromatography conditions: Chromatographic column: Acquity UPLC HSS C18 column (1.8 μm, 2.1 × 100 mm) Mobile phases: Phase A, 60 / 40 acetonitrile / water (5 mM ammonium acetate); Phase B, 90 / 10 isopropanol / acetonitrile (5 mM ammonium acetate). Flow rate: 0.3 mL / min Gradient elution procedure: as shown in Table 2.

[0060] Table 2. LC-MS / MS gradient elution program Mass spectrometry detection conditions: Heated ESI electrospray ionization source, positive ion ionization voltage 3.5 kV, negative ion ionization voltage -3.0 kV, ion source temperature 350℃, sheath gas flow rate 40 Arb, auxiliary gas flow rate 15 Arb, collision gas pressure 2.0 mTorr.

[0061] 2.3 Experimental Results Targeted lipidomics analysis of peripheral blood samples from patients with schizophrenia and healthy controls revealed significant changes in the relative levels of various lipid components in the peripheral blood of patients with schizophrenia compared to the healthy control group. The decreases in triglycerides (TG) and phospholipids were particularly pronounced (see...). Figure 1 ).

[0062] Further screening revealed the most significantly altered lipid metabolites in the plasma of schizophrenia patients, including TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2), and PE (18:1 / 18:2). The levels of these lipids in schizophrenia patients were significantly lower than in healthy controls (see [link to study]). Figure 2 (p<0.0001).

[0063] A diagnostic model for schizophrenia was constructed based on TG (40:0-FA16:0), PE (18:1 / 18:3), PE (18:2 / 18:2), and PE (18:1 / 18:2) and combinations of four lipids. Receiver operating characteristic (ROC) curves were used to analyze the model's diagnostic performance (see Table 3 and...). Figure 3The results showed that each individual lipid molecule exhibited good discriminative ability. Specifically, the area under the curve (AUC) for TG (40:0-FA16:0) was 0.831 (95% confidence interval: 0.752-0.894), with diagnostic sensitivity, specificity, and accuracy of 88.3%, 60.2%, and 71.4%, respectively; the AUC for PE (18:1 / 18:3) was 0.852 (95% confidence interval: 0.772-0.915), with diagnostic sensitivity, specificity, and accuracy of 81.9%, 69.1%, and 71.4%, respectively. The AUC of PE (18:2 / 18:2) was 0.864 (95% confidence interval: 0.791–0.923), with diagnostic sensitivity, specificity, and accuracy of 88.9%, 69.0%, and 76.8%, respectively. The AUC of PE (18:1 / 18:2) was 0.829 (95% confidence interval: 0.751–0.897), with diagnostic sensitivity, specificity, and accuracy of 79.3%, 67.1%, and 72.2%, respectively. Furthermore, after combining the above four lipids for modeling, the discrimination performance was significantly improved, with an AUC of 0.909 (95% confidence interval: 0.850–0.961), and diagnostic sensitivity, specificity, and accuracy of 95.6%, 82.2%, and 87.5%, respectively, showing superior discriminative ability. Furthermore, the model achieved a prediction precision of 79.0%, an F1 score of 0.862, a Matthews correlation coefficient (MCC) of 0.766, and a Cohen's Kappa coefficient of 0.751. All these metrics were calculated based on cross-validation predictions and the true labels. Specifically, prediction precision reflects the proportion of true positives among those predicted as positive; the F1 score is the harmonic mean of prediction precision and recall (i.e., sensitivity), used to comprehensively evaluate the model's performance under class imbalance; the MCC comprehensively considers true positives, true negatives, false positives, and false negatives, and is a robust metric for evaluating the overall performance of binary classification models; and the Cohen's Kappa coefficient measures the consistency between the predicted and true results and corrects for the impact of random consistency.

[0064] Table 3. Performance of the four lipid metabolite models In summary, the peripheral blood lipid biomarkers described in this invention can be applied to the clinical auxiliary diagnosis, disease risk screening, population stratification and related mechanism research of schizophrenia, and can serve as a key component in in vitro diagnostic products, biomarker programs or multi-omics joint analysis systems for schizophrenia, showing good clinical translation potential and industrial application prospects.

[0065] The specific embodiments of the present invention have been described in detail above, but they are merely examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions to the present invention are also within the scope of the present invention. Therefore, all equivalent changes and modifications made without departing from the spirit and scope of the present invention are covered within the scope of the present invention.

Claims

1. The use of any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2) and phosphatidylethanolamine (18:1 / 18:2) as biomarkers in the preparation of diagnostic products for schizophrenia.

2. The use of reagents for detecting any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the preparation of diagnostic products for schizophrenia.

3. The application according to claim 1 or 2, wherein the product comprises a reagent for detecting any one or more of the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the product includes reagents for detecting any one of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the product includes reagents for detecting triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the triglyceride (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), or phosphatidylethanolamine (18:1 / 18:2) samples are derived from peripheral blood; Preferably, the reagents used to detect the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2) and / or phosphatidylethanolamine (18:1 / 18:2) are the same or different.

4. The application according to claim 3, wherein, The reagents are reagents used for detection by chromatographic, spectroscopic, mass spectrometric, liquid chromatography-mass spectrometry, or chemical analysis methods; Preferably, the product comprises a C18 chromatographic column, mobile phase A being 60 / 40 acetonitrile / water containing 5 mM ammonium acetate, and mobile phase B being 90 / 10 isopropanol / acetonitrile containing 5 mM ammonium acetate; Preferably, the product is a reagent kit.

5. The application according to any one of claims 1-4, wherein, The diagnostic criteria are as follows: when the levels of any one or more of the following in the peripheral blood of the subject are significantly lower than those in healthy individuals, the subject is considered a suspected patient of schizophrenia. Preferably, the judgment criterion is as follows: when the levels of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the peripheral blood of the subject are all significantly lower than those in healthy individuals, the subject is judged to be a suspected patient of schizophrenia. Preferably, "significantly lower than" means p < 0.0001.

6. The application according to any one of claims 1-5, wherein, Triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2) and phosphatidylethanolamine (18:1 / 18:2) were used as biomarkers in combination.

7. A diagnostic kit for schizophrenia, wherein, The kit includes reagents for detecting any one or more of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the kit includes reagents for detecting any one of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the kit includes reagents for detecting triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2); Preferably, the triglyceride (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), or phosphatidylethanolamine (18:1 / 18:2) samples are derived from peripheral blood; Preferably, the reagents used to detect the triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2) and / or phosphatidylethanolamine (18:1 / 18:2) are the same or different.

8. The kit according to claim 7, wherein, The reagents are reagents used for detection by chromatographic, spectroscopic, mass spectrometric, liquid chromatography-mass spectrometry, or chemical analysis methods.

9. The kit according to claim 7 or 8, wherein, The kit includes a C18 column, mobile phase A being 60 / 40 acetonitrile / water containing 5 mM ammonium acetate, and mobile phase B being 90 / 10 isopropanol / acetonitrile containing 5 mM ammonium acetate.

10. The kit according to any one of claims 7-9, wherein, The diagnostic criteria for the kit are as follows: when the levels of any one or more of the following in the peripheral blood of the subject are significantly lower than those in healthy individuals, the subject is considered a suspected patient of schizophrenia. Preferably, the judgment criterion is as follows: when the levels of triglycerides (40:0-FA16:0), phosphatidylethanolamine (18:1 / 18:3), phosphatidylethanolamine (18:2 / 18:2), and phosphatidylethanolamine (18:1 / 18:2) in the peripheral blood of the subject are all significantly lower than those in healthy individuals, the subject is judged to be a suspected patient of schizophrenia. Preferably, "significantly lower than" means p < 0.0001.

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