CN114544790A - Application of reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in preparation of depression detection kit - Google Patents
Application of reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in preparation of depression detection kit Download PDFInfo
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- CN114544790A CN114544790A CN202011341741.8A CN202011341741A CN114544790A CN 114544790 A CN114544790 A CN 114544790A CN 202011341741 A CN202011341741 A CN 202011341741A CN 114544790 A CN114544790 A CN 114544790A
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- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 title claims abstract description 29
- 210000002381 plasma Anatomy 0.000 title claims abstract description 28
- CWRILEGKIAOYKP-SSDOTTSWSA-M [(2r)-3-acetyloxy-2-hydroxypropyl] 2-aminoethyl phosphate Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCCN CWRILEGKIAOYKP-SSDOTTSWSA-M 0.000 title claims abstract description 27
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 18
- 238000002360 preparation method Methods 0.000 title claims description 5
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 24
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 12
- 235000019253 formic acid Nutrition 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims description 5
- 238000004811 liquid chromatography Methods 0.000 claims description 4
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000003550 marker Substances 0.000 description 5
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 208000012902 Nervous system disease Diseases 0.000 description 2
- 208000025966 Neurological disease Diseases 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 238000002552 multiple reaction monitoring Methods 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 2
- 208000007848 Alcoholism Diseases 0.000 description 1
- 206010010144 Completed suicide Diseases 0.000 description 1
- 208000020401 Depressive disease Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102100037611 Lysophospholipase Human genes 0.000 description 1
- 208000019022 Mood disease Diseases 0.000 description 1
- 206010027951 Mood swings Diseases 0.000 description 1
- 108010058864 Phospholipases A2 Proteins 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002117 illicit drug Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/30—Psychoses; Psychiatry
- G01N2800/304—Mood disorders, e.g. bipolar, depression
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Abstract
The invention discloses application of a reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in preparing a depression detection kit, and belongs to the field of depression detection. The application of the invention is based on the difference of lysophosphatidylethanolamine (22:5) in the blood plasma of depression patients and normal people, and the difference is enough to accurately distinguish the depression patients from the normal people. The invention also discloses a detection kit based on the principle. The kit can realize quantifiable objective detection and has the advantages of sensitivity, rapidness and high reliability.
Description
Technical Field
The invention belongs to the field of depression detection, and particularly relates to application of a reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in preparation of a depression detection kit.
Background
Depression is a major type of mood disorder, with significant and persistent mood swings as the primary clinical feature. As one of the diseases with the highest disability rate in the world, the burden on families, healthcare systems and society by depression patients is enormous. It is estimated that depression symptoms are present in up to 50% of 80 million suicide worldwide per year. At present, the number of people suffering from depression in the world is more than 3.5 hundred million people, and more than 5400 million people in China suffer from depression.
The depression has obvious heredity, the heredity rate reaches 30-40 percent, and a plurality of genetic factors are involved. It is also affected by a number of non-genetic factors which increase the complexity of the pathogenesis of depression. The unclear pathogenesis causes that the diagnosis method for the disease always stays in subjective observation layers such as questionnaire scales and the like, lacks objective examination indexes, and has high operation difficulty and long diagnosis period. Therefore, it is urgent to develop a diagnostic method or apparatus based on objective indicators that can be quantified, which is sensitive, fast, and highly reliable.
Lysophosphatidylethanolamine naturally exists in animal cells or plant cells, and mainly exists in egg yolk or brain cells. Lysophosphatidylethanolamine is derived from phosphatidylethanolamine, which is a phospholipid containing 2 fatty acids, and when phospholipase a2 acts on phosphatidylethanolamine, one fatty acid is removed to produce lysophosphatidylethanolamine. Lysophosphatidylethanolamine (22:5) is one of lysophosphatidylethanolamines, wherein the numbers before the colon in the parentheses represent the number of C atoms, and the numbers after the colon represent the number of C ═ C double bonds. At present, the relationship between lysophosphatidylethanolamine (22:5) in blood and depression has not been reported.
Disclosure of Invention
The invention aims to solve the problems that: provided is a novel use of lysophosphatidylethanolamine (22:5) as a marker of depression.
The technical scheme of the invention is as follows:
use of a reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in the preparation of a depression detection kit.
Further, the reagent is used for a liquid chromatography-mass spectrometry combined method.
Further, the reagent is a reagent for a liquid chromatography method.
Further, the chromatographic column, 0.1% formic acid dissolved in pure water and 0.1% formic acid dissolved in acetonitrile are also included.
Further, the detection standard of the kit is as follows: when the plasma of the subject is significantly higher than that of the healthy population, the subject is judged to be likely to suffer from depression.
A depression detection kit comprising a reagent for detecting lysophosphatidylethanolamine (22:5) in plasma.
Further, the reagent is used for a liquid chromatography-mass spectrometry combined method.
Further, the reagent is a reagent for a liquid chromatography method.
Further, the chromatographic column, 0.1% formic acid dissolved in pure water and 0.1% formic acid dissolved in acetonitrile are also included.
Further, the detection standard of the kit is as follows: when the plasma of the subject is significantly higher than that of the healthy population, the subject is judged to be likely to suffer from depression.
The key point of the invention is that the level of human plasma lysophosphatidylethanolamine (22:5) is determined to be significantly related to depression, so that the risk of suffering from depression can be judged by detecting the level of human plasma lysophosphatidylethanolamine (22:5), and as for the specific means for detecting the human plasma lysophosphatidylethanolamine (22:5), the specific means can be any of various means disclosed in the prior art, and the liquid chromatography-mass spectrometry combined method is adopted for detection in the embodiment of the invention, but the method is not limited to the method.
The invention provides a novel depression detection marker and a novel depression detection kit, which can realize quantifiable objective detection of depression and have the advantages of sensitivity, rapidness and high reliability.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1: relative amounts of lysophosphatidylethanolamine (22:5) in depression and normal control plasma; CON, control; MDD, depression.
Detailed Description
Example 1 Depression detection kit and method of use thereof
1. Composition of the kit
Standard pure solution containing 12mg/ml plasma metabolism marker, 1 root C8BEH column (1.7 μm, 2.1X 100mm) and 1T3HSS column (1.8 μm, 2.1X 100mm), 0.1% formic acid dissolved in pure water, 0.1% formic acid dissolved in acetonitrile. Wherein the plasma metabolism marker is lysophosphatidylethanolamine (22: 5).
2. Application method
(a) Injecting the prepared plasma sample into a chromatographic column through an autosampler to separate metabolites respectively in a positive ionization mode and a negative ionization mode, wherein the specific chromatographic conditions are as follows: the mobile phase A is 0.1 percent formic acid dissolved in pure water; mobile phase B was acetonitrile-dissolved 0.1% formic acid. For positive mode, the gradient starts initially at 10% B, increases linearly to 40% B in 4 minutes after 1 minute, then increases to 100% B in 12 minutes and maintains for 5 minutes, then returns to the initial ratio equilibrium for about 3 minutes. In negative mode, the gradient starts at 100% a, increases linearly to 40% B after 1 minute, then increases to 100% B in 9 minutes, and remains for 4 minutes, then returns to the initial ratio equilibrium for about 4 minutes. The column temperature was 55 ℃.
(b) The separated metabolites were introduced into a liquid chromatography-mass spectrometer Shimadzu LC (30AD) -MS (TQ 8050) to obtain dynamic Multiple Reaction Monitoring (MRM) data for validation, the main parameters of the mass spectrum were the same as positive and negative modes: the flow rate of the heating gas is 10L/min; the flow rate of the dry gas flow is 10L/min; the flow rate of the atomization airflow is 3L/min; the DL temperature is 250 ℃; the temperature of the heating block is 400 ℃; the interface heater temperature was 300 ℃.
(c) Assessing whether the subject has depression based on the relative concentration of lysophosphatidylethanolamine (22: 5). A relative concentration >0.013 determines a high risk of suffering from depression.
The kit is designed based on the plasma metabolism marker provided by the invention, and can be used for accurately diagnosing and accurately evaluating depression patients.
To demonstrate the effectiveness of lysophosphatidylethanolamine (22:5) in evaluating depression, the following experimental examples are provided.
Experimental example 1 comparison of plasma lysophosphatidylethanolamine (22:5) concentration between patients with depression and normal controls
50 clinically diagnosed depression and normal controls were collected separately, and 5ml plasma samples were prepared for each collection. 100 samples were obtained from the first hospital affiliated with Chongqing medical university, and subjects in the depression group were excluded from past or present suffering from other neurological or psychiatric disorders, alcoholism or dependence on illicit drug use or pregnancy and were diagnosed as depression by the first hospital affiliated with Chongqing medical university using the DSM-IV-TR criteria and conducted the Hamilton Depression Scale to assess the severity of depression. The normal control group subjects exclude any past or present neurological disease, I or II axis disease or systemic medical disease. The study protocol fully met the ethical standards of human trials and was approved by the ethical committee of the university of Chongqing medical, with subjects informed prior to the trial and informed written consent.
The plasma sample is tested for plasma lysophosphatidylethanolamine (22:5) concentration by the kit and method of example 1, and the result is shown in fig. 1, wherein the plasma lysophosphatidylethanolamine (22:5) concentration of depression patients is significantly higher than that of the control (p)<0.001), FDR 0.0004, Log2FC 0.5718. According to fig. 1, the relative concentration of 0.013 can be used as a threshold for predicting depression, the relative concentration>0.013 the risk of the subject suffering from depression was determined to be high.
Note: FDR refers to the false discovery rate, an FDR of 0.0004 indicates an expected value of 0.0004 for the proportion of the number of false rejects (rejecting true (original) hypotheses) to the number of all rejected original hypotheses; FC refers to fold change.
The results of the experimental examples show that the plasma lysophosphatidylethanolamine (22:5) of the depression patients is obviously higher than that of the normal people, and the plasma lysophosphatidylethanolamine (22:5) concentration can be used for distinguishing the depression patients from the normal people.
In conclusion, the kit can be used for rapid auxiliary diagnosis of depression and has good application prospect.
Claims (10)
1. Use of a reagent for detecting lysophosphatidylethanolamine (22:5) in blood plasma in the preparation of a depression detection kit.
2. Use according to claim 1, characterized in that: the reagent is used in a liquid chromatography-mass spectrometry combined method.
3. Use according to claim 1, characterized in that: the reagent is used for a liquid chromatography method.
4. Use according to claim 2 or 3, characterized in that: the kit comprises T3HSS column, 0.1% formic acid dissolved in pure water as mobile phase a and 0.1% formic acid dissolved in acetonitrile as mobile phase B.
5. Use according to claim 1, characterized in that: the detection standard of the kit is as follows: when the plasma of the tested person is obviously higher than that of the healthy people, the person is judged to be a person with high risk of depression.
6. A depression detection kit, characterized in that: the kit comprises a reagent for detecting lysophosphatidylethanolamine (22:5) in plasma.
7. The kit of claim 6, wherein: the reagent is used in a liquid chromatography-mass spectrometry combined method.
8. The kit of claim 6, wherein: the reagent is used for a liquid chromatography method.
9. The kit of claim 7 or 8, wherein: the kit comprises T3HSS column, 0.1% formic acid dissolved in pure water as mobile phase a and 0.1% formic acid dissolved in acetonitrile as mobile phase B.
10. The kit of claim 6, wherein: the detection standard of the kit is as follows: when the plasma of the tested person is obviously higher than that of the healthy people, the person is judged to be a person with high risk of depression.
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| CN115219705A (en) * | 2022-07-14 | 2022-10-21 | 中国医学科学院北京协和医院 | Application of biomarker in Cushing syndrome diagnosis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115219705A (en) * | 2022-07-14 | 2022-10-21 | 中国医学科学院北京协和医院 | Application of biomarker in Cushing syndrome diagnosis |
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