Application of plasma lipid marker in early diagnosis kit for pheochromocytoma of adrenal gland

Abstract

The invention discloses application of a plasma lipid marker in an early diagnosis kit for pheochromocytoma of adrenal gland. The plasma lipid marker comprises LPC 18: 1, PC 6: 032: 2, PC 36: 5, TG 8: 08: 035: 1 and SM 35: 1. The ratio of O2 to Cer is 12: 0; o2 / 22: 5 is any one or more; the ratio of the LPC is 18: 1, the ratio of the PC is 6: 032: 2, the ratio of the PC is 36: 5, the ratio of the TG is 8: 08: 035: 1, and the ratio of the SM is 35: 1; the ratio of O2 to Cer is 12: 0; the accuracy of distinguishing a healthy subject from an adrenal pheochromocytoma patient by any one or combination of O2 / 22: 5 reaches 85% or above; therefore, the marker has high diagnosis accuracy on the pheochromocytoma of the adrenal gland and can be used for preparing a kit for early diagnosis of healthy people suffering from the pheochromocytoma of the adrenal gland.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of disease detection and diagnosis, and particularly relates to application of a plasma lipid marker in an early diagnosis kit for adrenal pheochromocytoma. BACKGROUND

[0002] Adrenal pheochromocytoma is a malignant tumor, which usually causes excessive secretion of catecholamine hormones, thereby causing various pathophysiological abnormalities of the human body. Although the incidence is very low, the possibility of pheochromocytoma should be considered in the diagnosis of hypertension, especially in the diagnosis of refractory hypertension, because 90% of pheochromocytomas can be cured, and once missed diagnosis, excessive catecholamines can cause fatal complications. Therefore, early diagnosis and appropriate treatment are very important for adrenal pheochromocytoma, which can avoid many potential complications and further development of the tumor. At present, the diagnosis of adrenal pheochromocytoma mainly adopts a combination of imaging examination and endocrine examination. Imaging examination mainly includes ultrasonography, CT, MRI and PET examination and other technologies, which can illustrate the occurrence and development of the disease through morphological characteristics, density and enhancement mode. CT is the preferred positioning examination method at present, and the diagnostic sensitivity reaches 93% to 100%, but the specificity is not high. For adrenal pheochromocytoma, there may be missed diagnosis, thereby causing many patients to need additional endocrine examination. Endocrine examination mainly judges the endocrine function of the tumor through biochemical analysis, mainly including methoxyepinephrine, methoxy-norepinephrine and other catecholamines and metabolites. However, these biochemical detection methods do not have good diagnostic accuracy, are not suitable for all populations, and the hormones are easily affected by external factors, and the detection results are unstable.

[0003] Lipidomics is a core branch of metabolomics and an important part of systems biology, which is a cross discipline that qualitatively identifies, quantitatively analyzes, and studies the structural characteristics, physical and chemical properties, metabolic rules of all lipid molecules in biological systems including cells, tissues, biological fluids, organisms, and the interactions between lipid molecules and other biological molecules and biological phenotypes on the whole level. In recent years, based on the advantages of high-throughput and high sensitivity of lipidomics technology, lipidomics has extremely important roles in new drug research and development, drug efficacy evaluation, early diagnosis of diseases and individualized treatment and other fields.

[0004] At present, the incidence of adrenal pheochromocytoma is increasing year by year, and we expect to achieve early screening and early diagnosis and timely treatment for the disease. Under this condition, we urgently need to research and develop markers with higher accuracy, higher sensitivity and higher specificity, and the plasma lipid marker diagnosis also shows obvious advantages. SUMMARY

[0005] The application aims to provide application of a plasma lipid marker in an early diagnosis kit for adrenal pheochromocytoma, so as to solve the problems of poor sensitivity and low accuracy in early diagnosis of adrenal pheochromocytoma.

[0006] To achieve the above-mentioned purpose, the application provides the following technical scheme.

[0007] The application provides application of a plasma lipid marker in preparation of an early diagnosis reagent or medicine for adrenal pheochromocytoma, wherein the plasma lipid marker comprises LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5.

[0008] The application also provides application of a plasma lipid marker in preparation of a kit for diagnosing early adrenal pheochromocytoma, wherein the kit comprises the plasma lipid marker, and the plasma lipid marker comprises LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5.

[0009] The application also provides a reagent or medicine for early diagnosis of adrenal pheochromocytoma, wherein the reagent or medicine comprises one or more plasma lipid markers in LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2, Cer 12:0;O2 / 22:5.

[0010] The application also provides a kit for early diagnosis of adrenal pheochromocytoma, wherein the kit comprises plasma lipid markers LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5.

[0011] Further, the diagnosis kit further comprises a standard sample of the plasma lipid marker.

[0012] Further, the diagnosis kit further comprises a standard sample of an internal standard compound.

[0013] Further, the internal standard compound comprises 1,2-dihexylphosphatidylcholine and [2H37]-nonadecanoic acid.

[0014] Further, the internal standard in the positive ion mode is 1,2-dihexylphosphatidylcholine, and the internal standard in the negative ion mode is [2H37]-nonadecanoic acid.

[0015] The application also provides a reagent for detecting plasma lipid markers, the plasma lipid markers including LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5.

[0016] The application also provides a use of a reagent for detecting plasma lipid markers in the preparation of a kit for diagnosing early adrenal pheochromocytoma, the plasma lipid markers including LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5.

[0017] Compared with the prior art, the application has the following beneficial effects:

[0018] The application first proposes the plasma lipid markers LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5 as early diagnosis markers of adrenal pheochromocytoma, and the plasma lipid markers LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2 and Cer 12:0;O2 / 22:5 can be used for preparing an early diagnosis kit for adrenal pheochromocytoma, the kit can accurately distinguish adrenal pheochromocytoma patients and reduce the misdiagnosis rate, wherein the accuracy of PC 6:0_32:2 and Cer 12:0;O2 / 22:5, PC 6:0_32:2 and TG 8:0_8:0_35:1, TG 8:0_8:0_35:1 and Cer 12:0;O2 / 22:5, PC 6:0_32:2 and SM 35:1;O2, and PC 6:0_32:2 and PC 36:5 in distinguishing healthy subjects from adrenal pheochromocytoma patients is more than 97%; and the operation is simple, the cost is low, and the detection can be completed in a short time to provide timely diagnosis basis for clinicians. Therefore, the above markers have high accuracy in the diagnosis of adrenal pheochromocytoma and can be used for preparing an early diagnosis kit for adrenal pheochromocytoma. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 The relative content of LPC 18:1 in the plasma samples of the healthy subjects and the adrenal pheochromocytoma patients;

[0020] Figure 2 The relative content of PC 6:0_32:2 in the plasma samples of the healthy subjects and the adrenal pheochromocytoma patients;

[0021] Figure 3 Relative amount of PC 36:5 in plasma samples of healthy subjects and patients with adrenal pheochromocytoma;

[0022] Figure 4 Relative amount of TG 8:0_8:0_35:1 in plasma samples of healthy subjects and patients with adrenal pheochromocytoma;

[0023] Figure 5 Relative amount of SM 35:1;O2 in plasma samples of healthy subjects and patients with adrenal pheochromocytoma;

[0024] Figure 6 Relative amount of Cer 12:0;O2 / 22:5 in plasma samples of healthy subjects and patients with adrenal pheochromocytoma;

[0025] Figure 7 ROC curve for LPC 18:1 alone to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0026] Figure 8 ROC curve for PC 6:0_32:2 alone to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0027] Figure 9 ROC curve for TG 8:0_8:0_35:1 alone to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0028] Figure 10 ROC curve for SM 35:1;O2 alone to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0029] Figure 11 ROC curve for Cer 12:0;O2 / 22:5 alone to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0030] Figure 12 ROC curve for LPC 18:1 and PC 6:0_32:2 in combination to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0031] Figure 13 ROC curve for LPC 18:1 and PC 36:5 in combination to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0032] Figure 14 ROC curve for LPC 18:1 and TG 8:0_8:0_35:1 in combination to diagnose healthy subjects and patients with adrenal pheochromocytoma;

[0033] Figure 15 ROC curve for the LPC 18:1 and Cer 12:0;02 / 22:5 combination to diagnose healthy subjects and pheochromocytoma patients;

[0034] Figure 16 ROC curve for the LPC 18:1 and Cer 12:0;02 / 22:5 combination to diagnose healthy subjects and pheochromocytoma patients;

[0035] Figure 17 ROC curve for the PC 6:0_32:2 and PC 36:5 combination to diagnose healthy subjects and pheochromocytoma patients;

[0036] Figure 18 ROC curve for the PC 6:0_32:2 and TG 8:0_8:0_35:1 combination to diagnose healthy subjects and pheochromocytoma patients;

[0037] Figure 19 ROC curve for the PC 6:0_32:2 and SM 35:1;02 combination to diagnose healthy subjects and pheochromocytoma patients;

[0038] Figure 20 ROC curve for the PC 6:0_32:2 and Cer 12:0;02 / 22:5 combination to diagnose healthy subjects and pheochromocytoma patients;

[0039] Figure 21 ROC curve for the PC 36:5 and TG 8:0_8:0_35:1 combination to diagnose healthy subjects and pheochromocytoma patients;

[0040] Figure 22 ROC curve for the PC 36:5 and SM 35:1;02 combination to diagnose healthy subjects and pheochromocytoma patients;

[0041] Figure 23 ROC curve for the PC 36:5 and Cer 12:0;02 / 22:5 combination to diagnose healthy subjects and pheochromocytoma patients;

[0042] Figure 24 ROC curve for the TG 8:0_8:0_35:1 and SM 35:1;02 combination to diagnose healthy subjects and pheochromocytoma patients;

[0043] Figure 25ROC curve of TG 8:0_8:0_35:1 and Cer 12:0;O2 / 22:5 combination for diagnosing the healthy subjects and the patients with adrenal pheochromocytoma;

[0044] Figure 26 ROC curve of SM 35:1;O2 and Cer 12:0;O2 / 22:5 combination for diagnosing the healthy subjects and the patients with adrenal pheochromocytoma. DETAILED DESCRIPTION

[0045] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.

[0046] Main reagents: methanol and acetonitrile (UPLC pure) were purchased from Merck Company in Germany, HPLC-grade formic acid and ammonium acetate were purchased from ROE Company in the United States; deionized water was prepared by a MIlli-Q ultrapure water system of Millipore Company in the United States; an internal standard 1,2-dihexylphosphatidylcholine (CAS: 34506-67-7 ) was purchased from Shanghai Zhenzhun Biological Technology Co., Ltd., and [2H37]-nonadecanoic acid (CAS: 646-30-0 ) was purchased from Shanghai Zhenzhun Biological Technology Co., Ltd.

[0047] All subjects obtained written informed consent before receiving the study.

[0048] The inclusion criteria of the healthy subjects and the patients with adrenal pheochromocytoma are as follows:

[0049] Healthy human subjects: no cardiovascular, respiratory, liver, kidney, gastrointestinal, endocrine, blood, mental, or nervous system diseases and disease history, no acute or chronic diseases. No drug allergy history, and the results of clinical laboratory tests were within the normal reference range at the time of screening.

[0050] Patients with adrenal pheochromocytoma: patients who have been diagnosed by pathological typing after preoperative examination and surgery;

[0051] Samples: collected in the First Affiliated Hospital of Zhejiang University, and 27 healthy subjects and 23 patients with adrenal pheochromocytoma were matched by age and gender according to strict screening and exclusion criteria.

[0052] Example 1

[0053] Diagnostic performance of target lipids for diagnosing the healthy subjects and the patients with adrenal pheochromocytoma

[0054] 1. Collection and storage of plasma samples

[0055] Peripheral blood was collected from patients with adrenal pheochromocytoma and healthy subjects in the morning on an empty stomach and collected into tubes containing an anticoagulant sodium citrate solution. All plasma samples were carefully aliquoted into sterile lyophilization tubes, labeled and stored at -80°C in a refrigerator for later use.

[0056] 2. Sample preparation

[0057] The plasma samples of step 1 were slowly thawed in a 4°C refrigerator for 30 min. 25 μL of plasma was taken from each sample in a 1.5 mL centrifuge tube, 200 μL of a methanol solution containing 2 μg / mL 1,2-dihexylphosphatidylcholine, 2 μg / mL [2H37]-nonadecanoic acid was added, and the mixture was mixed well by high-speed vortexing (>1600 rpm) for 1 min. After mixing, 750 μL of methyl tert-butyl ether was added, and the mixture was mixed well by high-speed vortexing (>1600 rpm) for 10 min. After the end of the vortexing, 200 μL of ultrapure water was added, and the mixture was mixed well by high-speed vortexing (>1600 rpm) for 5 min. The mixture was left to stand at room temperature for 10 min and then centrifuged at 13000 rpm at 4°C for 10 min in a low-temperature high-speed centrifuge. After centrifugation, 600 μL of the supernatant was taken and divided equally into two 1.5 mL centrifuge tubes for nitrogen blowing. After the supernatant was blown dry, 100 μL of a mixed solvent of acetonitrile / isopropyl alcohol / water (65:35:5) was added for re-dissolution, and the mixture was mixed well by high-speed vortexing (>1600 rpm) for 10 min. After mixing, the mixture was centrifuged at 13000 rpm at 4°C for 5 min in a low-temperature high-speed centrifuge. After centrifugation, 70 μL of the supernatant was taken for injection. In addition, 10 μL of each sample was mixed as a quality control sample. The mixture was capped and placed in a liquid chromatograph-mass spectrometer for preparation for positive and negative ion detection. The ratio of the peak area of each target lipid substance LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2, and Cer 12:0;O2 / 22:5 to the peak area of the internal standard was taken as the relative content of the metabolite.

[0058] 3. Determination of the relative content of target lipid substances in plasma by UPLC-Q / TOF-MS

[0059] Instrument: Agilent 1290 ultra-performance liquid chromatography coupled with Agilent 6545 quadrupole-time-of-flight mass spectrometer. Chromatographic conditions: Column: Waters BEH C8 (100 mm x 2.1 mm I.D., 1.7 μm); mobile phase: acetonitrile-isopropanol (8:2, v:v) (B) - water (A) solution (containing 10 mM ammonium acetate formic acid); flow rate: 0.3 mL / min; column temperature: 55 °C, autosampler maintained at 10 °C, injection volume 2 μL. Elution gradient as follows: 0-2.5 min, 40-30% A; 2.5-8 min, 30%-15% A; 8-12 min, 15% A; 12-14 min, 15%-10% A; 14-17 min, 10-1% A; 17-22 min, 1% A; 22-23 min, 40% A. Run time 23 min, post run time 5 min.

[0060] Mass spectrometric conditions: Collision voltage 120 V, cone hole voltage 65 V, Oct 1 RF Vpp voltage 750 V; drying gas temperature 350 °C, drying gas flow rate 12 L / min, nebulizer pressure 35 psi, sheath gas temperature 350 °C, sheath gas flow rate 11 L / min, both drying gas and cone hole gas are high purity nitrogen; capillary voltage 3000 V, nozzle voltage 1500 V; ion source temperature 120 °C.

[0061] 4. Data processing method

[0062] Lipid metabolite data were analyzed using unsupervised principal component analysis (PCA) and supervised orthogonal partial least squares discriminant analysis (OPLS-DA) to obtain clustering information and differentially expressed metabolites. Univariate statistical analysis among metabolites was performed using the false discovery rate (FDR) to correct for p-values. Variables simultaneously satisfying VIP (Variable Importance in Projection) > 1 and P < 0.05 were selected as differentially expressed metabolites. The primary and secondary spectra of differentially expressed metabolite peaks were identified and compared using MS-DIAL and Agilent PCDLManager B.07.00 databases. For the selected differentially expressed metabolites, the Least Absolute Shrinkage and Selection Operator (LASSO) model was used to evaluate the contribution of eight metabolites to pheochromocytoma and normal samples. The Area Under Curve (AUC) values ​​for the diagnosis of pheochromocytoma were evaluated using Receiver Operating Characteristic Curve (ROC) curves.

[0063] 5. Experimental Results

[0064] (1) Differences in the levels of target lipids in the plasma of healthy subjects and patients with pheochromocytoma of the adrenal gland.

[0065] like Figures 1-5 As shown, compared with healthy subjects, the relative levels of target lipids LPC 18:1, PC 6:0.32:2, PC 36:5, TG 8:0.8:0.35:1, and SM 35:1;O2 were significantly downregulated in plasma samples from patients with adrenal pheochromocytoma. Figure 6 As shown, the relative levels of the target lipid Cer 12:0;O2 / 22:5 were significantly upregulated in plasma samples from patients with adrenal pheochromocytoma.

[0066] (2) ROC curves of target lipids alone in differentiating healthy subjects from patients with pheochromocytoma of the adrenal gland

[0067] The ROC curve results for distinguishing between healthy subjects and patients with adrenal pheochromocytoma are shown in Table 2 and... Figures 7-11 As shown, Figure 7 As shown, the area under the ROC curve for LPC 18:1 in differentiating healthy subjects from patients with adrenal pheochromocytoma was 0.848. Figure 8The area under the ROC curve for PC 6:0_32:2, used to differentiate between healthy subjects and patients with adrenal pheochromocytoma, was 0.906, as shown in Table 2. The area under the ROC curve for PC 36:5, used to differentiate between healthy subjects and patients with adrenal pheochromocytoma, was 0.861, as shown in Table 2. Figure 9 As shown, the area under the ROC curve for TG 8:0_8:0_35:1 in differentiating healthy subjects from patients with adrenal pheochromocytoma is 0.922. Figure 10 As shown, the area under the ROC curve for SM 35:1;O2 in distinguishing healthy subjects from patients with adrenal pheochromocytoma was 0.871. Figure 11 As shown, the area under the ROC curve for Cer 12:0;O2 / 22:5, which differentiates healthy subjects from patients with adrenal pheochromocytoma, is 0.938.

[0068] The principle of ROC curve evaluation method:

[0069] Basic evaluation indicators for diagnostic tests include sensitivity and specificity, while comprehensive evaluation indicators include the Youden index, ROC, and AUC. For the evaluation of diagnostic tests, it is first necessary to determine the true group of the sample being tested using the gold standard. For the disease group (equivalent to the pheochromocytoma group in this invention) and the healthy group (equivalent to the healthy subject group in this invention) determined by the gold standard, the results of the diagnostic test can be categorized as follows:

[0070] Positive (True Positive, TP); the diagnostic test result is positive (consistent with the gold standard result);

[0071] Negative (True Negative, TN); the diagnostic test result was negative (consistent with the gold standard result);

[0072] False positive (FP): A positive result in a diagnostic test (inconsistent with the gold standard result);

[0073] False negative (FN): A diagnostic test result is negative (inconsistent with the gold standard result).

[0074] This can be represented by the following table 1:

[0075] Table 1. Diagnostic test results

[0076]

[0077] The sensitivity of a diagnostic test = A / (A+C); the specificity of a diagnostic test = D / (B+D). Sensitivity and specificity determine the diagnostic sensitivity and specificity of a test relative to the gold standard. High sensitivity means fewer cases will be diagnosed as negative, resulting in a low rate of missed diagnoses; high specificity means fewer healthy cases will be diagnosed as positive, resulting in a low rate of false positives.

[0078] The ROC curve is a curve plotted based on the aforementioned sensitivity and specificity. Using the possible diagnostic cutoff values ​​in the diagnostic test as diagnostic points, the corresponding sensitivity and specificity are calculated according to the table above. Then, with sensitivity as the ordinate and 1-specificity as the abscissa, the sensitivity and specificity points at each diagnostic point are plotted on a coordinate graph. Connecting the coordinate points yields a smooth curve, which is the ROC curve. The more and denser the diagnostic points are, the smoother the resulting ROC curve will be.

[0079] The ROC curve uses each test result as a possible diagnostic cutoff value, and the area under the curve (AUC) indicates the accuracy of the diagnostic test. The AUC is widely accepted as an inherent accuracy indicator for evaluating the validity of diagnostic tests. An AUC of 0.5 indicates no diagnostic significance; an AUC between 0.5 and 0.7 indicates low diagnostic accuracy; an AUC between 0.7 and 0.9 indicates moderate diagnostic accuracy; and an AUC greater than 0.9 indicates high diagnostic accuracy. Higher AUC values ​​indicate higher diagnostic accuracy.

[0080] Table 2. ROC curves of target lipid substances alone distinguishing healthy subjects from patients with pheochromocytoma.

[0081]

[0082] ROC curve results showed that these six lipids—LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2, and Cer 12:0;O2 / 22:5—had high accuracy in distinguishing between healthy subjects and patients with adrenal pheochromocytoma when used alone, with TG 8:0_8:0_35:1 showing the highest accuracy (AUC 0.922).

[0083] (3) ROC curves for the diagnosis and differentiation between healthy subjects and patients with pheochromocytoma of the adrenal gland by pairwise combination of target lipid substances.

[0084] The ROC curve results for pairwise combinations of the six lipids LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:5 to diagnose and differentiate between healthy subjects and patients with adrenal pheochromocytoma are as follows: Figures 12-26 As shown in Table 3. Figure 12 The area under the ROC curve for the combination of LPC 18:1 and PC 6:0_32:2 in differentiating healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.928. Figure 13 As shown, the area under the ROC curve for the combination of LPC 18:1 and PC 36:5 in distinguishing healthy subjects from patients with adrenocortical pheochromocytoma was 0.875. Figure 14 As shown, the area under the ROC curve for the combination of LPC18:1 and TG 8:0_8:0_35:1 in differentiating healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.943. Figure 15 As shown, the area under the ROC curve for the LPC 18:1 and SM 35:1;O2 combination in distinguishing healthy subjects from patients with adrenocortical pheochromocytoma was 0.911. Figure 16 As shown, the area under the ROC curve for the combination of LPC 18:1 and Cer 12:0;O2 / 22:5 in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.935. Figure 17 As shown, the area under the ROC curve for the combination of PC 6:0_32:2 and PC 36:5 in differentiating healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.970. Figure 18 As shown, the area under the ROC curve for the combination of PC 6:0_32:2 and TG 8:0_8:0_35:1 in distinguishing healthy subjects from patients with adrenocortical pheochromocytoma was 0.988. Figure 19 As shown, the area under the ROC curve for the PC 6:0_32:2 and SM35:1;O2 combination in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.972. Figure 20 As shown, the area under the ROC curve for the combination of PC 6:0_32:2 and Cer 12:0;O2 / 22:5 in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.990. Figure 21 As shown, the area under the ROC curve for the combination of PC 36:5 and TG 8:0_8:0_35:1 in differentiating healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.962. Figure 22 As shown, the area under the ROC curve for the PC36:5 and SM 35:1;O2 combination in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.943.Figure 23 As shown, the area under the ROC curve for the combination of PC 36:5 and Cer 12:0;O2 / 22:5 in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.962. Figure 24 As shown, the area under the ROC curve for the TG 8:0_8:0_35:1 and SM 35:1;O2 combination in distinguishing healthy subjects from patients with adrenocortical pheochromocytoma was 0.950. Figure 25 As shown, the area under the ROC curve for the combination of TG 8:0_8:0_35:1 and Cer 12:0;O2 / 22:5 in distinguishing healthy subjects from patients with adrenocortical pheochromocytoma was 0.978. Figure 26 As shown, the area under the ROC curve for the combination of SM 35:1;O and Cer 12:0;O2 / 22:5 in distinguishing healthy subjects from patients with pheochromocytoma of the adrenal cortex was 0.960.

[0085] Table 3. ROC curves for pairwise combinations of target lipid substances to differentiate between healthy subjects and patients with pheochromocytoma.

[0086]

[0087] ROC curve results showed that pairwise combinations of six lipids—LPC 18:1, PC 6:0_32:2, PC 36:5, TG 8:0_8:0_35:1, SM 35:1;O2, and Cer 12:0;O2 / 22:5—had high accuracy in distinguishing between healthy subjects and patients with adrenal pheochromocytoma. Specifically, the combinations of PC 6:0_32:2 and Cer 12:0;O2 / 22:5, PC 6:0_32:2 and TG 8:0_8:0_35:1, TG 8:0_8:0_35:1 and Cer 12:0;O2 / 22:5, PC 6:0_32:2 and SM 35:1;O2, and PC 6:0_32:2 and PC... The 36:5 combination has extremely high accuracy (AUC of 0.990, 0.988, 0.978, 0.972, and 0.970), and has extremely high diagnostic value.

[0088] Example 2

[0089] Diagnostic kit for differentiating healthy subjects from patients with adrenal pheochromocytoma

[0090] A diagnostic kit for early diagnosis of pheochromocytoma in healthy individuals contains standards of diagnostic markers LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:5, and internal standard compounds 1,2-dihexamethylenetetramine and [2H37]-nonadecanoic acid.

[0091] Example 3

[0092] Diagnostic kit for differentiating healthy subjects from patients with adrenal pheochromocytoma

[0093] A diagnostic kit for early detection of pheochromocytoma in healthy individuals, comprising standards containing diagnostic biomarkers, wherein the diagnostic biomarkers are any one of LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1;O2 and Cer12:0;O2 / 22:5, and internal standard compounds including standards of 1,2-dihexamethylenetetramine and [2H37]-nonadecanoic acid.

[0094] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0095] It should be noted that the above content merely illustrates the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. For those skilled in the art, various improvements and modifications can be made without departing from the principle of the present invention, and all such improvements and modifications fall within the scope of protection of the claims of the present invention.

Claims

1. The application of a plasma lipid marker in the preparation of early diagnostic reagents or drugs for adrenal pheochromocytoma, characterized in that, The plasma lipid markers include LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:

5.

2. The application of a plasma lipid marker in the preparation of a kit for diagnosing early-stage adrenal pheochromocytoma, characterized in that, The plasma lipid markers include LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:

5.

3. A reagent or drug for the early diagnosis of adrenal pheochromocytoma, characterized in that, The reagents or drugs include one or more plasma lipid markers selected from LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2, Cer 12:0; O2 / 22:

5.

4. A kit for early diagnosis of adrenal pheochromocytoma, characterized in that, The kit includes one or more plasma lipid markers selected from LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2, Cer 12:0; O2 / 22:

5.

5. The kit for early diagnosis of adrenal pheochromocytoma according to claim 4, characterized in that, The kit also includes standards for the plasma lipid markers.

6. The kit for early diagnosis of adrenal pheochromocytoma according to claim 4, characterized in that, The kit also includes a standard of internal standard compound.

7. A kit for early diagnosis of adrenal pheochromocytoma according to claim 6, characterized in that, The internal standard compounds include 1,2-dihexylphospholipin and [2H37]-nonadecanoic acid.

8. A reagent for detecting plasma lipid markers, characterized in that, The plasma lipid markers include LPC18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:

5.

9. The application of a reagent for detecting plasma lipid markers in the preparation of a kit for diagnosing early-stage adrenal pheochromocytoma, characterized in that, The plasma lipid markers include LPC 18:1, PC 6:0-32:2, PC 36:5, TG 8:0-8:0-35:1, SM 35:1; O2 and Cer 12:0; O2 / 22:5.

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