Application of membrane lipidome in preparation of teenager depression diagnosis product

Abstract

The invention provides an application of a membrane lipid group in preparation of a product for diagnosing juvenile depression, and particularly relates to an erythrocyte membrane lipid group which is selected from the following groups: ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesteryl ester (ChE). The membrane lipidome is stable, is not easily interfered by dietary habits and other factors, can avoid subjective concealing, and provides a more objective, real and stable method for diagnosing juvenile depression.

Description

Technical Field

[0001] The present invention relates to the field of disease diagnosis, and in particular to use of a membrane lipidome in preparing a product for diagnosing adolescent depression. Background Art

[0002] Depression is one of the most common mental disorders in adolescence, with a lifetime prevalence of up to 19% by the end of adolescence. The main characteristics of depression include persistent negative emotions, decreased motivation and interest. Depression continues to impair adolescents' psychological functioning and quality of life, becoming the fourth largest disease burden among young people aged 10-24 years.

[0003] At present, the International Classification of Diseases (ICD), the American Diagnostic and Statistical Manual of Mental Disorders (DSM), and the Chinese Classification of Mental Disorders (CCMD) are mainly used to diagnose depression at home and abroad. However, diagnostic tools such as ICD and DSM are mainly based on the patient's clinical symptoms, and the operation is complicated and time-consuming. Therefore, they require experienced psychiatric specialists to operate, which is not conducive to the early and efficient screening and rapid diagnosis of adolescent depression. In addition, since adolescent depression patients often show unexplained physical symptoms, eating disorders, anxiety, refusal to go to school, poor academic performance, drug abuse or behavioral problems, adolescent depression is often easily missed or misdiagnosed. At the same time, the extremely high heterogeneity of adolescent depression patients also limits their accurate and effective diagnosis to a certain extent.

[0004] Some standardized assessment tools are used to help identify adolescent depression, including the Beck Depression Inventory (BDI) and the Children's Depression Inventory (CDI). However, questionnaires and scales used for adolescent depression are unstructured diagnoses and are highly subjective. Some studies have reported that metabolites in plasma or serum may serve as diagnostic markers for adolescent depression, but serum and plasma metabolites are easily disturbed by short-term lifestyle habits such as diet and exercise.

[0005] Therefore, exploring objective and stable biomarkers has profound clinical and practical significance for promoting efficient screening and diagnosis of adolescent depression. Summary of the invention

[0006] The purpose of the present invention is to provide an objective and stable biomarker for diagnosing adolescent depression.

[0007] In a first aspect of the present invention, a method for determining adolescent depression in a non-diagnostic and non-therapeutic manner is provided, the method comprising the steps of: detecting the expression level of membrane lipids separated in a red blood cell membrane sample of a subject to be tested, thereby determining whether the subject to be tested is a patient with adolescent depression; and

[0008] The membrane lipids are selected from the group consisting of ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE).

[0009] In another preferred embodiment, the red blood cell membrane lipids are selected from the following group: Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1).

[0010] In another preferred embodiment, the method further comprises: after measuring the concentration of the membrane lipid in the sample, using the above concentration to calculate the P value, thereby reaching a judgment conclusion.

[0011] In another preferred embodiment, the P value is calculated using the following logic model:

[0012]

[0013] The concentration of Cer (d19:1_24:1) is c1;

[0014] The concentration of Cer (d16:1_16:1) is c2;

[0015] The concentration of PS (22:4_18:2) is c3;

[0016] The concentration of PC (22:5_18:2) is c4;

[0017] The concentration of PC (18:2_20:4) is c5;

[0018] The concentration of PC (18:1e_20:3) is c6;

[0019] The concentration of TG (18:4_18:1_18:2) is c7;

[0020] The concentration of ChE(18:1) is c8.

[0021] In another preferred embodiment, the detection standard of the product is: when the subject's P value>0.396, it is determined to be a patient with adolescent depression.

[0022] In another preferred embodiment, the concentration of the membrane lipid in the sample is tested by liquid chromatography-mass spectrometry.

[0023] In another preferred embodiment, the chromatographic conditions of the liquid chromatography-mass spectrometry method are: the chromatographic column is ACQUITYUPLC BEH C18, phase A is a mixed solution of acetonitrile and water with ammonium formate and formic acid added, and phase B is a mixed solution of isopropanol acetonitrile with ammonium formate and formic acid added, and gradient elution is performed.

[0024] In another preferred embodiment, the phase A is a mixed solution of acetonitrile and water to which 5-20 mM ammonium formate and 0.01-0.2% vv formic acid are added.

[0025] In another preferred embodiment, the phase A is a mixed solution of acetonitrile water (volume ratio of 2-10:2-6) added with 5-20 mM ammonium formate and 0.01-0.2% vv formic acid.

[0026] In another preferred embodiment, the phase B is a mixed solution of isopropanol and acetonitrile to which 5-20 mM ammonium formate and 0.01-0.2% vv formic acid are added.

[0027] In another preferred embodiment, the phase B is a mixed solution of isopropanol acetonitrile (volume ratio of 6-15:0.5-2) added with 5-20 mM ammonium formate and 0.01-0.2% vv formic acid.

[0028] In another preferred embodiment, the method further comprises the step of: using a pretreatment unit to process the blood sample before performing the membrane lipid group concentration determination.

[0029] In another preferred embodiment, the processing method comprises the steps of collecting red blood cells, extracting red blood cell membranes, and then extracting lipids in the red blood cell membranes.

[0030] In another preferred embodiment, the processing method comprises the steps of:

[0031] (1) Collecting red blood cells: Centrifuge the blood and take out the red blood cells in the lower layer;

[0032] (2) Extracting red blood cell membranes: Red blood cells were treated with 5-50 volumes of Tris-HCl (5-15 mM, pH = 7-8) at 1-10° C. to fully swell the red blood cells;

[0033] (3) Extraction of lipids: Add extraction solution (methanol: chloroform volume ratio of 0.1-2:1-5) to the red blood cell membrane precipitate, mix well and extract at 20-60°C for 1-5 hours. Then, vacuum centrifuge and dry at 15-30°C for 0.1-10 hours, dissolve the lipid extract in a re-solution (methylene chloride: isopropanol: methanol volume ratio of 0.1-2:0.1-2:1-5), mix well and centrifuge, and take the supernatant.

[0034] In a second aspect of the present invention, a kit for detecting depression in adolescents is provided, the kit comprising:

[0035] A sample pretreatment reagent, wherein the sample pretreatment reagent is used to extract red blood cell membrane lipids from a blood sample;

[0036] The erythrocyte membrane lipidome detection reagent is used to detect the expression level of membrane lipids separated in the erythrocyte membrane sample of the object to be detected.

[0037] In another preferred embodiment, the sample pretreatment reagent comprises:

[0038] Red blood cell membrane extraction reagent; preferably, the cell membrane extraction reagent is Tris-HCl;

[0039] Lipid extraction reagent; preferably, the lipid extraction reagent comprises a lipid extract (methanol: chloroform volume ratio of 0.1-2:1-5) and a lipid reconstitution solution (dichloromethane: isopropanol: methanol volume ratio of 0.1-2:0.1-2:1-5).

[0040] In another preferred embodiment, the kit further comprises an instruction manual, and the instruction manual records:

[0041] Detecting the expression level of membrane lipids separated from the red blood cell membrane sample of the test subject, thereby determining whether the test subject is a patient with adolescent depression; and

[0042] The membrane lipids are selected from the group consisting of ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE).

[0043] In another preferred embodiment, the red blood cell membrane lipids are selected from the following group: Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1).

[0044] In another preferred embodiment, the method further comprises: after measuring the concentration of the membrane lipid in the sample, using the above concentration to calculate the P value, thereby reaching a judgment conclusion.

[0045] In another preferred embodiment, the P value is calculated using the following logic model:

[0046]

[0047] The concentration of Cer (d19:1_24:1) is c1;

[0048] The concentration of Cer (d16:1_16:1) is c2;

[0049] The concentration of PS (22:4_18:2) is c3;

[0050] The concentration of PC (22:5_18:2) is c4;

[0051] The concentration of PC (18:2_20:4) is c5;

[0052] The concentration of PC (18:1e_20:3) is c6;

[0053] The concentration of TG (18:4_18:1_18:2) is c7;

[0054] The concentration of ChE(18:1) is c8.

[0055] In the third aspect of the present invention, a combination of membrane lipid markers in red blood cell membrane samples is provided for use in constructing a system for diagnosing adolescent depression; the membrane lipid markers are selected from the following group: ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE).

[0056] In another preferred embodiment, the red blood cell membrane lipid marker is selected from the following group: Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1).

[0057] In another preferred embodiment, the application further comprises: calculating the P value using the concentration of the membrane lipid marker in the sample to obtain a calculation result.

[0058] In another preferred embodiment, the P value is calculated using the following logic model:

[0059]

[0060] The concentration of Cer (d19:1_24:1) is c1;

[0061] The concentration of Cer (d16:1_16:1) is c2;

[0062] The concentration of PS (22:4_18:2) is c3;

[0063] The concentration of PC (22:5_18:2) is c4;

[0064] The concentration of PC (18:2_20:4) is c5;

[0065] The concentration of PC (18:1e_20:3) is c6;

[0066] The concentration of TG (18:4_18:1_18:2) is c7;

[0067] The concentration of ChE(18:1) is c8.

[0068] In another preferred example, when the returned P value>0.396, the patient is diagnosed with adolescent depression.

[0069] In another preferred embodiment, the system comprises a high performance liquid chromatography-mass spectrometry device.

[0070] In another preferred embodiment, the concentration of the membrane lipid marker in the sample is tested by liquid chromatography-mass spectrometry.

[0071] In another preferred embodiment, the chromatographic conditions of the liquid chromatography-mass spectrometry method are: the chromatographic column is ACQUITYUPLC BEH C18, phase A is a mixed solution of acetonitrile and water with ammonium formate and formic acid added, and phase B is a mixed solution of isopropanol acetonitrile with ammonium formate and formic acid added, and gradient elution is performed.

[0072] In another preferred embodiment, the phase A is a mixed solution of acetonitrile and water to which 5-20 mM ammonium formate and 0.01-0.2% vv formic acid are added.

[0073] In another preferred embodiment, the phase A is a mixed solution of acetonitrile water (volume ratio of 2-10:2-6) added with 5-20 mM ammonium formate and 0.01-0.2% vv formic acid.

[0074] In another preferred embodiment, the phase B is a mixed solution of isopropanol and acetonitrile to which 5-20 mM ammonium formate and 0.01-0.2% vv formic acid are added.

[0075] In another preferred embodiment, the phase B is a mixed solution of isopropanol acetonitrile (volume ratio of 6-15:0.5-2) added with 5-20 mM ammonium formate and 0.01-0.2% vv formic acid.

[0076] In another preferred embodiment, the method further comprises the step of: using a pretreatment unit to process the blood sample before performing the determination of the membrane lipid marker combination concentration.

[0077] In another preferred embodiment, the processing method comprises the steps of collecting red blood cells, extracting red blood cell membranes, and then extracting lipids in the red blood cell membranes.

[0078] In another preferred embodiment, the processing method comprises the steps of:

[0079] (1) Collecting red blood cells: Centrifuge the blood and take out the red blood cells in the lower layer;

[0080] (2) Extracting red blood cell membranes: Red blood cells were treated with 5-50 volumes of Tris-HCl (5-15 mM, pH = 7-8) at 1-10° C. to fully swell the red blood cells;

[0081] (3) Extraction of lipids: Add extraction solution (methanol: chloroform volume ratio of 0.1-2:1-5) to the red blood cell membrane precipitate, mix well and extract at 20-60°C for 1-5 hours. Then, vacuum centrifuge and dry at 15-30°C for 0.1-10 hours, dissolve the lipid extract in a re-solution (methylene chloride: isopropanol: methanol volume ratio of 0.1-2:0.1-2:1-5), mix well and centrifuge, and take the supernatant.

[0082] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1 is the standard curve of five lipid standards.

[0084] Figure 2 The diagnostic efficacy of eight red blood cell membrane lipids for adolescent depression. DETAILED DESCRIPTION

[0085] After extensive and in-depth research, the inventors have provided a use of a membrane lipid group in the preparation of a product for diagnosing adolescent depression through a large number of screenings and tests. The membrane lipid group is a red blood cell membrane lipid selected from the following group: ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE). After collecting red blood cells from the subject's venous blood, the membrane lipids are separated and extracted, and the red blood cell membrane lipids in the subject's blood are detected by ultra-high performance chromatography-mass spectrometry to obtain the peak area of ​​each lipid. The lipid concentration is then calculated with the aid of the standard curve of the standard. And a P value formula is obtained with lipid concentration as an independent variable, wherein the P value and are the risk values ​​of the subject suffering from adolescent depression, and the larger the P value, the higher the risk level. The membrane lipid is stable and is not easily disturbed by confounding factors such as short-term diet, exercise and drugs. It is closely related to the pathological mechanism of adolescent depression patients, truly reflects the health status of the subject, and avoids subjective deception of the subject compared to clinical interviews and scales, providing a more objective, true and stable method for diagnosing adolescent depression. The present invention was completed on this basis.

[0086] the term

[0087] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0088] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0089] As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0090] As used herein, the term "room temperature" or "normal temperature" refers to a temperature of 4-40°C, preferably, 25±5°C.

[0091] The main advantages of the present invention include:

[0092] 1. The membrane lipidome of the present invention can objectively and truly reflect the health status of the subject. Compared with clinical interviews and scales, it avoids the subjective deception of the subject, may reduce misdiagnosis and missed diagnosis, and is more accurate.

[0093] 2. The membrane lipidome described in the present invention is a combination of multiple membrane lipids, and has higher true accuracy, precision and AUC value than a single lipid.

[0094] 3. The concentration of membrane lipids described in the present invention is more stable than plasma / serum lipids / metabolites and is not easily interfered by confounding factors such as short-term diet, exercise and drugs. It is a relatively more stable biomarker.

[0095] 4. The membrane lipidome extraction method described in the present invention is based on lipidomics and is targeted to extract red blood cell membrane lipids. The method is simple and has good effects.

[0096] 4. The present invention provides a method for quantifying red blood cell membrane lipids, which performs absolute quantification of individual red blood cell membrane lipids through an external standard curve, which can more truly reflect the content of lipids in the body compared with the peak area. There is no previous report on the quantification of individual red blood cell membrane lipids in adolescent patients with depression.

[0097] The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0098] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0099] Example 1

[0100] The present invention detects the red blood cell membrane lipidome by ultra-high performance liquid chromatography-mass spectrometry to quantify the level of specific lipids. The operation steps are described in detail as follows:

[0101] 1) Collecting red blood cells: Collect 2 mL of venous blood into an EDTA anticoagulant tube, invert several times to mix, centrifuge at 1600 g for 10 minutes, and remove the red blood cells in the lower layer for later use.

[0102] 2) Extraction of erythrocyte membranes: erythrocytes were treated with 20 volumes of Tris-HCl (10 mM, pH 7.4) at 4°C overnight to fully swell the erythrocytes, and then washed three times with PBS.

[0103] 3) Extraction of lipids: Add extraction solution (methanol: chloroform volume ratio of 1:2) to the red blood cell membrane precipitate, vortex mix for 5 minutes, transfer to a glass injection bottle, and extract in a 40°C constant temperature box for 2 hours. Then, vacuum centrifuge and dry at room temperature for 1 hour, dissolve the lipid extract with a re-solution (dichloromethane: isopropanol: methanol volume ratio of 1:1:2), mix well, transfer to a 1.5 mL centrifuge tube, and take the supernatant for detection.

[0104] 4) Preparation of lipid standards: Ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE) were quantified by Cer (d18:1 / 18:0), PS (17:0 / 17:0), PC (19:0 / 19:0), TG (17:0 / 17:1 / 17:0) D5 and cholesterol standards, respectively, and prepared into the following concentration gradients with complex solutions (Table 1).

[0105] Table 1 Standard concentration gradient

[0106]

[0107] 5) On-machine detection: Ultra-high performance liquid chromatography-quadrupole orbital trap mass spectrometry (Vanquish UHPLC / QExactive plus) was used to detect erythrocyte membrane lipids. The ACQUITY UPLC BEH C18 column (100×2.1mm, 1.7μm, Waters) was set at a column temperature of 55°C. Mobile phase A: acetonitrile: water volume ratio 6:4, plus 10mM ammonium formate and 0.1% formic acid, mobile phase B: isopropanol: acetonitrile volume ratio 9:1, plus 10mM ammonium formate and 0.1% formic acid; the flow rate was 400μL / min. The chromatographic elution gradient was from 95 / 5 to 0 / 100 within 17min. The detection was performed in positive and negative ion modes, respectively, with the following operating parameters: spray voltage of 3.8 kV (positive ion mode) and 3.0 kV (negative ion mode); heated capillary temperature of 320 °C; data-dependent acquisition mode, 1 full scan, 6 MS / MS scans; full scan range: 150-2000 amu; full scan resolution: 70000, automatic gain control (AGC) <1×10 6 , scanning time 0.1s; MS / MS scanning resolution: 17500, AGC <5×10 5 , scanning time 0.05s. Concentration curve of standard was established.

[0108] 5) Lipid quantification: Extract the peak areas of Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1), and quantify them using standards of their respective categories. Cer (d19:1_24:1) was quantified using the Cer (d18:1 / 18:0) standard curve in positive ion mode, Cer (d16:1_16:1) was quantified using the Cer (d18:1 / 18:0) standard curve in negative ion mode, PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1) were quantified using their respective standards, and the concentrations of these eight lipids were denoted as c1 to c8, respectively.

[0109] The concentrations of these eight lipids were used as independent variables to establish a logistic regression model, and formula ① is as follows:

[0110]

[0111] According to the above method, the concentrations of these eight lipids in the red blood cell membrane of each subject were detected and calculated. Substituting them into formula ①, the resulting P is the risk value of the subject suffering from adolescent depression. The larger the P value, the higher the risk level.

[0112] According to the specific application scenario, an appropriate P value threshold can be selected. Preferably, when P>0.396, the subject is judged to have adolescent depression ( Figure 2 ), and the AUC is 0.971.

[0113] At the same time, the control group used the concentration of a single lipid as the independent variable to establish a logistic regression model and obtain a curve chart, in which the AUC of Cer (d19:1_24:1) was 0.808, the AUC of Cer (d16:1_16:1) was 0.813, the AUC of PS (22:4_18:2) was 0.806, the AUC of PC (22:5_18:2) was 0.797, the concentration AUC of PC (18:2_20:4) was 0.806, the AUC of PC (18:1e_20:3) was 0.825, the AUC of TG (18:4_18:1_18:2) was 0.801, and the AUC of ChE (18:1) was 0.748. It can be seen that the AUC of any single lipid is lower than 0.971 (lipid group), so the diagnostic potential of the combination of these eight red blood cell membrane lipids is significantly better than the effect of any single lipid.

[0114] Example 2 Evaluation of the detection effect of membrane lipidome

[0115] Based on the data of 81 adolescent patients with depression and 67 healthy controls, we calculated the identification results of the membrane lipid group for adolescent patients with depression under the condition of P>0.396, as shown in Table 2.

[0116] Table 2 Summary of statistical results

[0117]

[0118] Based on the data in the above table, the consistency rate between membrane lipid group and clinical diagnosis results was calculated through the following aspects:

[0119] 1. Sensitivity, specificity and accuracy

[0120] Sensitivity is the ability of the membrane lipid group to correctly identify adolescent patients with depression. The higher the value, the more effective the membrane lipid group is in detecting adolescent patients with depression. The calculation method is: Sensitivity = number of true positives / (number of true positives + number of false negatives).

[0121] In the present invention, the sensitivity of the membrane lipid group = 77 / 81 = 95.1%, which is high.

[0122] Specificity measures the ability of the membrane lipidome to correctly identify healthy controls. A higher value indicates that the membrane lipidome is more effective in excluding healthy controls. The calculation method is: Specificity = number of true negatives / (number of true negatives + number of false positives).

[0123] In the present invention, the specificity of the membrane lipidome = 58 / 67 = 86.6%, which is high.

[0124] Accuracy measures the proportion of samples correctly classified by the membrane lipidome diagnostic model as a whole, and is calculated as: Accuracy = (number of true positives + number of true negatives) / number of samples.

[0125] In the present invention, the accuracy of the membrane lipidome = (58 + 77) / 148 = 91.2%, which is highly accurate.

[0126] 2. Kappa consistency analysis

[0127] The Kappa coefficient is a statistic used to evaluate the consistency of classifiers in classification problems, especially to evaluate the consistency between classification results and actual conditions. When the Kappa coefficient is greater than 0.8, it means that the membrane lipidome and clinical diagnosis have very good consistency, 0.6 < Kappa coefficient ≤ 0.8, it is considered that the consistency of the two methods is high, 0.4 < Kappa coefficient ≤ 0.6, it has moderate consistency, 0.2 < Kappa coefficient ≤ 0.4, it is generally consistent, and Kappa coefficient ≤ 0.2 is considered to be very poor consistency.

[0128] The Kappa coefficient calculation formula is as follows:

[0129] Kappa = (Po-Pe) / (1-Pe)

[0130] Among them, Po is the observation consistency and Pe is the observation consistency.

[0131] The calculation methods of Po and Pe are as follows:

[0132] Po = (58 + 77) / 148 = 91.2%

[0133] Pe=(62*67+86*81) / (148*148)=50.8%

[0134] Therefore, Kappa coefficient = (91.2% - 50.8%) / (1-50.8%) = 0.821, indicating that the membrane lipidome has a very high consistency with the clinical diagnosis.

[0135] 3.Matthews correlation coefficient

[0136] The Matthews correlation coefficient is one of the indicators used to evaluate the performance of a binary classification model. It takes into account four different prediction results: true positive, true negative, false positive, and false negative, and provides a performance evaluation that is balanced across all categories. The Matthews correlation coefficient ranges from -1 to +1, where +1 indicates complete prediction agreement, 0 indicates that the prediction is no better than random prediction, and -1 indicates complete prediction inconsistency. The closer it is to 1, the better the classification performance of the model. The calculation method is:

[0137]

[0138] Therefore, the Matthews correlation coefficient between the membrane lipidome and clinical diagnosis was 0.824, indicating that the membrane lipidome diagnostic model performed well in identifying adolescent depression patients.

[0139] In summary, the membrane lipidome provided by the present invention shows high sensitivity, high specificity, and high accuracy in diagnosing adolescent depression, and has extremely high consistency with clinical diagnostic results and excellent classification performance.

[0140] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A method for non-diagnostic and non-therapeutic determination of adolescent depression, characterized in that: The method comprises the steps of: detecting the expression level of membrane lipids separated in the red blood cell membrane sample of the subject to be tested, thereby determining whether the subject to be tested is a patient with adolescent depression; and The membrane lipids are selected from the group consisting of ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE).

2. The method according to claim 1, characterized in that The red blood cell membrane lipids are selected from the following group: Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1).

3. The method according to claim 1, characterized in that The method further comprises: after measuring the concentration of the membrane lipid in the sample, using the above concentration to calculate the P value, thereby obtaining a judgment conclusion.

4. The method according to claim 3, characterized in that The P value is calculated using the logistic model shown in the following formula ①: The concentration of Cer (d19:1_24:1) is c1; The concentration of Cer (d16:1_16:1) is c2; The concentration of PS (22:4_18:2) is c3; The concentration of PC (22:5_18:2) is c4; The concentration of PC (18:2_20:4) is c5; The concentration of PC (18:1e_20:3) is c6; The concentration of TG (18:4_18:1_18:2) is c7; The concentration of ChE(18:1) is c8.

5. The method according to claim 3, characterized in that The detection standard of the product is: when the subject's P value>0.396, he or she is judged to be a patient with adolescent depression.

6. The method according to claim 3, characterized in that The concentration of the membrane lipids in the sample was tested by liquid chromatography-mass spectrometry.

7. A test kit for adolescent depression, characterized in that: The kit comprises: A sample pretreatment reagent, wherein the sample pretreatment reagent is used to extract red blood cell membrane lipids from a blood sample; The erythrocyte membrane lipidome detection reagent is used to detect the expression level of membrane lipids separated in the erythrocyte membrane sample of the object to be tested.

8. The detection kit according to claim 7, characterized in that The sample pretreatment reagent comprises: Red blood cell membrane extraction reagent; preferably, the cell membrane extraction reagent is Tris-HCl; Lipid extraction reagent; preferably, the lipid extraction reagent comprises a lipid extract (methanol: chloroform volume ratio of 0.1-2:1-5) and a lipid reconstitution solution (dichloromethane: isopropanol: methanol volume ratio of 0.1-2:0.1-2:1-5).

9. The detection kit according to claim 7, characterized in that The kit also includes instructions, and the instructions record: Detecting the expression level of membrane lipids separated in the red blood cell membrane sample of the test subject, thereby determining whether the test subject is a patient with adolescent depression; and The membrane lipids are selected from the group consisting of ceramide (Cer), phosphatidylserine (PS), phosphatidylcholine (PC), triglyceride (TG) and cholesterol ester (ChE).

10. The detection kit according to claim 9, characterized in that The red blood cell membrane lipids are selected from the following group: Cer (d19:1_24:1), Cer (d16:1_16:1), PS (22:4_18:2), PC (22:5_18:2), PC (18:2_20:4), PC (18:1e_20:3), TG (18:4_18:1_18:2) and ChE (18:1).

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