CN105445408A - Metabolite marker for diagnosing and distinguishing coronary atherosclerosis and stable angina pectoris - Google Patents
Metabolite marker for diagnosing and distinguishing coronary atherosclerosis and stable angina pectoris Download PDFInfo
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Abstract
The invention discloses a metabolite marker for diagnosing and distinguishing coronary atherosclerosis and stable angina pectoris. The metabolite marker includes one or more of phosphorylcholine, palmitoylethanolamine, phytosphingosine, phosphatidylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidyl inositol (20:4/0:0). When the single metabolite marker is used for diagnosing and distinguishing a patient suffering from the stable angina pectoris and a patient suffering from the coronary atherosclerosis, the ROC areas under the curve (AUC) are greater than 0.7, and the marker has clinical diagnostic significance. When the markers are combined for diagnosis, the AUC is further improved with the increase of the combination number; when all eight markers are combined, the AUC is the highest and is up to 0.985, and the sensitivity and specificity are 97.4% and 98.0% respectively under the condition of an optimal cutoff value. The metabolite marker can accurately diagnose and distinguish the coronary atherosclerosis and the stable angina pectoris and is high in accuracy and good in sensitivity and specificity.
Description
Technical field
The invention belongs to biochemical field, relate to the metabolic markers of diagnosis of coronary heart disease somatotype, being specifically related to one group for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis.
Background technology
Coronary heart disease is also called ischemic heart disease, the artery relating to supply myocardial blood occurs atherosis, namely coronary atherosclerosis causes the narrow or Mottling formation of lumen of vessels even to break, block completely, cause myocardial ischemia, anoxic or necrosis and cause heart ischemia disease, thus causing a series of serious cardiovascular event of angina pectoris, myocardial infarction etc. clinically.Coronary heart disease is the primary killers of human health, the features such as the tool incidence of disease is high, disability rate is high, recurrence rate is high, case fatality rate is high, complication is many, has become the principal disease threatening our people's health.
At present, based on pathophysiological mechanism, coronary heart disease is divided into chronic stable coronary heart disease (i.e. stable angina cordis) and acute coronary syndrome; Acute coronary syndrome is further divided into again unstable angina pectoris and acute myocardial infarction AMI.Coronary heart disease is from without to having, from light to heavily generally including following developing stage: coronary artery is normal → and coronary atherosclerosis → stable angina cordis → unstable angina pectoris → acute myocardial infarction AMI.
Coronary atherosclerosis is the Etiological of coronary heart disease, is the early stage illness of coronary heart disease.Coronary atherosclerosis is a kind of common Progressive symmetric erythrokeratodermia arterial disease; pathology mainly involves medium sized muscle layer artery; endarterium lipidosis; smooth muscle cell proliferation; form limitation patch, arterial wall can be made hardening, and time serious, plaque rupture causes thrombus, embolism, hemorrhage; the luminal part or entirely shut of getting involved, clinical manifestation is the generation of atherosclerotic blood vessel complication.Before Earlier atherosclerotic lesion can occur in 10 years old; pathology causes arteriarctia need experience for 20 to 30 years; early stage without clinical symptoms, be not easily found and pay attention to, therefore to the early prevention of coronary atherosclerosis with diagnose the generation that effectively can prevent coronary heart disease.
Coronarography can be made stenosis coronarius and being judged accurately, goldstandard [coronarography goldstandard and the comparative studies of clinical routine diagnostics coronary heart disease otherness of diagnosis of coronary heart disease, Shu Rongwen etc., naval medicine magazine 04 phase in 2015].As the goldstandard of diagnosis of coronary heart disease, coronary angiography can only find hemadostewnosis degree, and it or a kind of get involved measurement means, need intervene operation, and diagnosis is expensive.On the other hand, doctor also needs according to making last diagnostic to the check result such as cardiogram, cardiogram, Treadmill Exercise Test, CT of patient, due to the appearance that doctor's subjectivity judges, patient describes the situation such as unclear, the diagnosis of coronary heart disease is still existed to larger mistaken diagnosis and fails to pinpoint a disease in diagnosis, and this is very large to the influence prognosis of patient.In order to improve the quality of life of patient, reduce the life threat that is subject to of patient, we need badly development a kind of have diagnosis high, economical, without the diagnostic method of invasive, the characteristic such as easy and simple to handle.
Metabolism group be postgraduate's object endogenous metabolism material entirety and with internal cause and external cause change science, be an important component part of systems biology.It can carry out fast body fluids such as blood and urine and without invasive analysis, the difference of being composed by metabolism obtains the metabolic markers that can indicate various biochemical reaction.Analytical technology conventional at present comprises nuclear magnetic resonance (NMR), mass spectrum (LC-MS/GC-MS) etc.LC-MS/GC-MS has and requires the features such as low, highly sensitive, wide dynamic range to sample preparation, can be used for detecting the metabolin that in sample, concentration differs greatly, thus becomes in the research of metabolism group and apply increasing technology platform.Plasma analysis is a kind of methods for the diagnosis of diseases commonly used clinically, is widely adopted because of its easy, quick, economic and advantage of relative noninvasive.
Not yet someone uses blood plasma metabolite level to carry out diagnosis typing to coronary heart disease at present.The application blood plasma metabolism group level difference of finding coronary artery normal person and coronary atherosclerosis patients and different somatotype Patients Plasma with Coronary Heart Disease metabolin makes a definite diagnosis coronary heart disease and to carry out somatotype significant fast in early days for clinical.
Summary of the invention
In order to overcome the deficiencies in the prior art, the present invention first object is to provide one group for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, and this group metabolic markers is present in blood plasma simultaneously, can analyze mensuration simultaneously; The second object of the present invention is that providing a kind of can analyze the method detecting described metabolic markers delicately; The third object of the present invention is to provide a kind of detection kit based on described metabolic markers, distinguishes coronary atherosclerosis and stable angina cordis for diagnosing, and improves diagnosis convenience, promotes diagnostic method standardization.
Above-mentioned purpose is achieved by following technical solution:
One group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises one or more metabolic markers as described below: phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0).
Lysophosphatidyl choline (16:0), i.e. 1-palmitoyl-glycerol-3-phosphatid ylcholine.
Lysophosphatidyl choline (18:2), i.e. 1-sub-oleoyl-glycerol-3-phosphatid ylcholine.
Phosphatidylinositols (20:4/0:0), i.e. 1-arachidonic acyl glycerophosphatide acyl inositol.
Further, described one group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises the metabolic markers described in any three.
Further, described one group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises the metabolic markers described in any four.
Further, described one group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises the metabolic markers described in any five.
Further, described one group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises the metabolic markers described in any six.
Further, described one group, for diagnosing the metabolic markers of differentiation coronary atherosclerosis and stable angina cordis, comprises the metabolic markers described in whole eight.
Further, described metabolic markers derives from blood plasma.
Described in a kind of qualitative or quantitative test for diagnosing the method for the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis to be: adopt LC-MS and/or gas chromatography mass spectrometry to carry out qualitative or quantitative test to described metabolic markers.Liquid matter gentle quality detection limit is low, highly sensitive, can analyze delicately to detect metabolic markers in biological specimen and quantitative to it.
For diagnosing the detection kit distinguishing coronary atherosclerosis and stable angina cordis, comprise the standard items of described metabolic markers, described standard items are chemical monomer or the potpourri of each metabolic markers.Use standard items can carry out qualitative and quantitative analysis rapidly and accurately to the metabolic markers in biological specimen.Kit contributes to realizing standardized testing, improves and detects convenience and reappearance.
Further, described detection kit also comprise dissolve described standard items solvent and/or Extraction and enrichment described in the solvent of metabolic markers.
Advantage of the present invention:
(1) metabolic markers provided by the invention can be diagnosed exactly and distinguish coronary atherosclerosis and stable angina cordis.In ROC evaluating method for curve, ROC area under curve AUC, when being greater than 0.5, more close to 1, illustrates that diagnosis effect is better.AUC has lower accuracy 0.5 ~ 0.7 time, and AUC has certain accuracy 0.7 ~ 0.9 time, has high accuracy when AUC is more than 0.9.Empirical tests, metabolic markers provided by the invention single for diagnose distinguish coronary atherosclerosis and stable angina cordis time, AUC is all more than 0.7; During multiple use in conjunction, AUC is than single closer to 1, and diagnosis effect is better; During eight use in conjunction, AUC is closest to 1, and it is best that effect is distinguished in diagnosis.
(2) method of the described metabolic markers of analysis detection provided by the invention is highly sensitive, and result accurately and reliably.
(3) detection kit provided by the invention may be used for diagnosis differentiation coronary atherosclerosis and stable angina cordis, improves diagnosis convenience, promotes diagnostic method standardization.
Embodiment
Essentiality content of the present invention is further illustrated below in conjunction with embodiment.What the instrument used or reagent did not elaborate is conventional instrument and reagent; Not specifically described experimental working technique is conventional practices known to a person of ordinary skill in the art.
Embodiment 1: between coronary atherosclerosis patients and patients with stable angina pectoris, the screening of blood plasma difference metabolin characterizes
One, object and method
1, Specimen origin
After obtaining patient's agreement, collect the peripheric venous blood blood plasma of Jiangsu Prov. People's Hospital in September, 2010 ~ 2015 routine coronary atherosclerosis patients in year June 480,280 routine patients with stable angina pectoris and 350 routine Healthy Peoples, all patients or health confirm through coronary angiography per capita.Age of Healthy People, sex and coronary atherosclerosis patients, patients with stable angina pectoris match.All coronary atherosclerosis patients, patients with stable angina pectoris and health have normal cardiopulmonary liver kidney and hematopoiesis function per capita.
Blood sampling time is fasted conditions in early morning.
2, main agents
Acetonitrile and formic acid (UPLC is pure) are purchased from ROE company of the U.S.; Chromatogram rank methyl alcohol and chloroform are purchased from Hanbon Sci. & Tech. Co., Ltd.; Chlorination methoxamine and N-methyl-N-(trimethyl silane) trifluoroacetamide (containing 1% trimethyl chlorosilane) are purchased from Sigma-Aldrich; Deionized water is prepared by the MIlli-Q ultrapure water system of U.S. Mi Libo (Millipore) company; Standard items comprise phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0), are purchased from U.S. Sigma-Aldrich.
3, the screening of blood plasma difference metabolin characterizes
3.1UPLC-Q/TOF-MS screens sign
3.1.1 sample preparation
Response phase method is utilized to carry out Extraction solvent optimization: with the peak number under mass spectrum ESI+ and ESI-detecting pattern and total peak area for factor investigates different solvents (acetonitrile, methyl alcohol, ethanol, chloroform, water) to the Extraction and enrichment efficiency of metabolin in blood plasma.Experiment data measured is carried out multivariable analysis, utilizes importance factor in PLS model (variableimportancetoprojection, VIP value) to reflect the importance that variable responds model.The VIP value of acetonitrile, methyl alcohol, ethanol, chloroform, water is followed successively by 1.503,0.802,0.651,0.688 and 0.987, and the extraction efficiency of acetonitrile is the highest, therefore selects acetonitrile as the Extraction solvent of plasma sample.
Sample preparation: get 100 μ L blood plasma in 1.5mL centrifuge tube, add 400 μ L acetonitriles, vortex mixed after 30 seconds, 13000rpm × 10min centrifugal (4 DEG C), get 200 μ L supernatants in 1.5mL centrifuge tube, at room temperature dry up with Nitrogen evaporator, 20% acetonitrile solution of residue obtained use 300 μ L dissolves, and to obtain final product.
3.1.2 test condition and parameter
UPLC-Q/TOF-MS condition:
Chromatographic resolution adopts Ultra Performance Liquid Chromatography (UPLC, Agilent1290, USA).Chromatographic column is WatersBEHC18 post (100mm × 2.1mm, 1.7 μm), column temperature 25 DEG C, and Sample Room temperature is room temperature, sample size 2 μ L.Positive and negative ion pattern flowing phase composition is all A is volumetric concentration 0.1% aqueous formic acid, and B is volumetric concentration 0.1% formic acid acetonitrile solution.Condition of gradient elution: 0 ~ 1min is 0 ~ 30%B phase, and in 2min, B is phase linear is increased to 60%, and 3 ~ 8min linear change, to 90%B phase, is then linearly increased to 100%B phase at 8 ~ 9min and keeps 1min; Flow velocity 0.3mL/min, after post, efflux directly imports mass spectrometer system detection without shunting.
Mass spectrophotometry adopts level Four bar-flight time mass spectrum (Agilent6530Q-TOF/MS, USA).With electric spray ion source (ESI) positive and negative ion mode detection; Dry gas flow velocity is 7L/min, and dry gas temperature is 300 DEG C, and dry gas and taper hole gas are high pure nitrogen; Ion source temperature 100 DEG C, under positive ion and negative ion mode, capillary voltage is 3000V, and collision voltage is 100V; Employing full scan pattern is per second carries out three secondary data collections, quality of scanning scope: m/z100-1000 dalton.
3.2GC-Q/MS screens sign
3.2.1 sample preparation
Get 200 μ L blood plasma in 1.5mL centrifuge tube, add the interior mark of 2-isopropylmalate acid solution of 50 μ L1mg/mL, vortex mixing in 20 seconds, add 400 μ L methyl alcohol, the mixed solution (ratio is 2.5: 1: 1) of chloroform and water, then jolting 30min (1200rpm) on the metal bath of 70 DEG C, 16000g × 5min centrifugal (4 DEG C), get 500 μ L supernatants in 1.5mL centrifuge tube, add 500 μ L distilled water, vortex mixes, then 16000g × 5min centrifugal (4 DEG C), get 500 μ L supernatants in 1.5mL centrifuge tube, at room temperature dry up with Nitrogen evaporator, the methoxamine pyridine solution of residue obtained use 80 μ L dissolves, oximate 8h under 50 DEG C of conditions, add the trimethyl silicon based trifluoroacetamide of 60 μ LN-methyl-N-, derivatization 2h under 70 DEG C of conditions, obtain.
3.2.2 test condition and parameter
GC-Q/MS condition: U.S. Agilent7890B-5977A gas chromatograph-mass spectrometer (GCMS).Chromatographic column HP-5MS capillary column (30.0m × 0.25mm, capillary thickness 0.25 μm); Carrier gas is high-purity helium, flow velocity 1.0mL/min; Sample size 2 μ L; Temperature programme: 80 DEG C of constant temperature 2min, 80 DEG C-300 DEG C (5 DEG C/min) constant temperature 6min; Do not shunt, injector temperature 300 DEG C; Interface temperature 300 DEG C; Ion source temperature 200 DEG C; Electron energy 50eV; Solvent delay 3min; Adopt full scan pattern, quality of scanning scope: m/z30-600 dalton.
4, data processing and analysis
Data importing SIMCA software (version13.0.2, Umetrics) obtained by UPLC-Q/TOF-MS and GC-Q/MS carries out multivariate statistical analysis.By setting up OPLS-DA (orthogonal ginsenoside) model, find the metabolin of metabolic profile contribution between coronary atherosclerosis patients and patients with stable angina pectoris comparatively large (VIP > 1.0 and p < 0.01).
Carried out the retrieval of the structure of matter by databases such as HMDB (http://www.hmdb.ca/) and Metline (http://metlin.scripps.edu/), the structure of the above-mentioned difference metabolin of MS/MS collection of illustrative plates Preliminary Identification of accurate molecular weight and the mass spectrum gained provided in database is provided.Eventually through purchase standard items, with the molecular weight of standard items, chromatographic retention and the comparison of corresponding multistage MS fragmentation pattern, the structure of confirmation difference metabolin.
Two, result
Screening symbolizes 8 difference metabolins, is respectively: phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0).
Compared with coronary sclerosis patient, the expression of above-mentioned 8 difference metabolins in patients with stable angina pectoris blood plasma is all lowered.Quantitatively detected by standard items, compared with coronary sclerosis patient, the expression of above-mentioned difference metabolin in patients with stable angina pectoris blood plasma lowers 0.7 ~ 0.8 times.As can be seen here, the expression of above-mentioned 8 difference metabolins in patients with stable angina pectoris and coronary sclerosis patients blood plasma is obviously different, may be used for diagnosis and distinguishes coronary atherosclerosis and stable angina cordis.
Embodiment 2: build ROC curve and verify that 8 difference metabolins are for diagnosing the ability distinguishing coronary atherosclerosis and stable angina cordis
Adopting receiver operating curves (ROC) method to verify, judging that it is for diagnosing the ability distinguishing patients with stable angina pectoris and coronary atherosclerosis patients by the expression of difference metabolin in patients with stable angina pectoris and coronary atherosclerosis patients blood plasma.Result shows, these 8 difference metabolins of phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0) are single for diagnosing the ability distinguishing patients with stable angina pectoris and coronary atherosclerosis patients stronger, ROC area under curve (AUC) is all greater than 0.7, has clinical diagnosis meaning; When combining for diagnosing, along with the increase of associating number, AUC improves further, and the highest during 8 whole associatings, AUC reaches 0.985, and under best cutoff value, sensitivity and specificity are respectively 97.4% and 98.0%.Single and any 2 ~ 7 Combining diagnosis the results are shown in Table 1 ~ 3.
Coronary atherosclerosis patients and patients with stable angina pectoris are distinguished in the single difference metabolin diagnosis of table 1
| Single difference metabolin | AUC | Sensitivity | Specificity |
| Phosphocholine | 0.867 | 86.6％ | 87.8％ |
| Palmitylethanolamide | 0.854 | 83.3％ | 84.5％ |
| Phytosphingosine | 0.832 | 81.1％ | 82.3％ |
| Phosphatid ylcholine | 0.795 | 77.4％ | 78.6％ |
| Ethyl chenodeoxycholic acid | 0.779 | 75.8％ | 77.0％ |
| Lysophosphatidyl choline (16:0) | 0.758 | 73.7％ | 74.9％ |
| Lysophosphatidyl choline (18:2) | 0.733 | 71.2％ | 72.4％ |
| Phosphatidylinositols (20:4/0:0) | 0.711 | 70.1％ | 71.3％ |
Table 2 two difference metabolin Combining diagnosis distinguish coronary atherosclerosis patients and patients with stable angina pectoris
Any three ~ seven difference metabolin Combining diagnosis of table 3 distinguish coronary atherosclerosis patients and patients with stable angina pectoris
| Associating number | AUC | Sensitivity | Specificity |
| Three | ≥0.907 | ≥90.8％ | ≥90.2％ |
| Four | ≥0.919 | ≥91.3％ | ≥92.0％ |
| Five | ≥0.923 | ≥93.6％ | ≥93.4％ |
| Six | ≥0.946 | ≥95.2％ | ≥94.8％ |
| Seven | ≥0.968 | ≥96.3％ | ≥96.5％ |
As can be seen from Table 1, these 8 difference metabolins are single, and for diagnosing the ability distinguishing coronary atherosclerosis patients and patients with stable angina pectoris comparatively strong, AUC is all greater than 0.7, and sensitivity is higher, specificity is comparatively strong, has clinical diagnosis meaning; As can be seen from Table 2, when these 8 difference metabolins are combined between two for diagnosing, AUC than single for diagnosing time higher, highly sensitive, specificity is higher, has clinical diagnosis meaning; As can be seen from Table 3, when combining for diagnosing with 3 ~ 7 in these 8 difference metabolins, AUC improves further, highly sensitive, high specificity, has clinical diagnosis meaning.
Therefore, these 8 difference metabolins can as the metabolic markers for diagnosing differentiation coronary atherosclerosis and stable angina cordis.
Embodiment 3: the preparation of detection kit
Prepared detection kit based on metabolic markers provided by the invention, this kit comprises following composition:
The standard items of metabolic markers: comprise phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0), each standard items encapsulate respectively;
Blood plasma metabolin Extraction solvent: 100% acetonitrile and 20% acetonitrile solution (for UPLC-Q/TOF-MS sample preparation); Ratio be 2.5: 1: 1 methyl alcohol, the mixed solution of chloroform and water, methoxamine pyridine and the trimethyl silicon based trifluoroacetamide of N-methyl-N-(for GC-Q/MS sample preparation); During UPLC-Q/TOF-MS screening characterizes, 20% acetonitrile solution can be used as the solvent dissolving standard items; During GC-Q/MS screening characterizes, with the method preparation standard product solution of blood plasma metabolin Extraction solvent by sample preparation;
Interior mark: 2-isopropylmolic acid.
Certainly, during design detection kit, do not need the standard items comprising above-mentioned 8 metabolic markers completely, only can use wherein several combination.These standard items can encapsulate separately, also can make potpourri encapsulation.
This kit designs based on metabolic markers provided by the invention, may be used for diagnosis and distinguishes coronary atherosclerosis patients and patients with stable angina pectoris.
In sum, the present invention effectively overcomes deficiency of the prior art, and tool high industrial utilization.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.Those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the protection domain of technical solution of the present invention.
Claims (10)
1. one group for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that, comprise one or more metabolic markers as described below: phosphocholine, Palmitylethanolamide, phytosphingosine, phosphatid ylcholine, ethyl chenodeoxycholic acid, lysophosphatidyl choline (16:0), lysophosphatidyl choline (18:2) and phosphatidylinositols (20:4/0:0).
2. according to claim 1 for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: comprise the metabolic markers described in any three.
3. according to claim 1 for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: comprise the metabolic markers described in any four.
4. according to claim 1 for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: comprise the metabolic markers described in any five.
5. according to claim 1 for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: comprise the metabolic markers described in any six.
6. according to claim 1 for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: comprise the metabolic markers described in whole eight.
7., according to the arbitrary described metabolic markers for diagnosing differentiation coronary atherosclerosis and stable angina cordis of claim 1 ~ 6, it is characterized in that: described metabolic markers derives from blood plasma.
8. the arbitrary described method for diagnosing the metabolic markers distinguishing coronary atherosclerosis and stable angina cordis of qualitative or quantitative test claim 1 ~ 6, is characterized in that: adopt LC-MS and/or gas chromatography mass spectrometry to carry out qualitative or quantitative test to described metabolic markers.
9. one kind for diagnosing the detection kit distinguishing coronary atherosclerosis and stable angina cordis, it is characterized in that: the standard items comprising the arbitrary described metabolic markers of claim 1 ~ 6, described standard items are chemical monomer or the potpourri of each metabolic markers.
10. detection kit according to claim 9, is characterized in that: also comprise dissolve described standard items solvent and/or Extraction and enrichment described in the solvent of metabolic markers.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102901789A (en) * | 2012-09-21 | 2013-01-30 | 中国药科大学 | Determination method of serum metabolic marker for early diagnosis of diabetic nephropathy. |
| CN103154742A (en) * | 2010-06-20 | 2013-06-12 | 佐拉生物科学公司 | Lipidomic biomarkers for identification of high-risk coronary artery disease patients |
| JP2015065979A (en) * | 2013-09-26 | 2015-04-13 | テルモ株式会社 | Guide wire |
| KR20150059644A (en) * | 2015-05-01 | 2015-06-01 | 학교법인 동아학숙 | composition for diagnosing coronary artery disease comprising CD31 or fragment thereof |
| US20150272999A1 (en) * | 2014-03-31 | 2015-10-01 | Banner Health | Retrograde coronary vein delivery of stem cells |
| US20150376704A1 (en) * | 2009-12-09 | 2015-12-31 | Cleveland Heartlab, Inc. | Biomarker assay for diagnosis and classification of cardiovascular disease |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1789806A2 (en) * | 2004-09-13 | 2007-05-30 | Lipomics Technologies, Inc. | Metabolite markers for weight management |
| CN101802620A (en) * | 2007-02-22 | 2010-08-11 | 特提斯生物科学公司 | Metabolic markers of diabetic conditions and methods of use thereof |
| CN101939651A (en) * | 2007-10-10 | 2011-01-05 | 保函医药公司 | Methods for detecting major adverse cardiovascular and cerebrovascular events |
| CN102421358A (en) * | 2009-03-10 | 2012-04-18 | 杜克大学 | Predicting coronary artery disease and risk of cardiovascular events |
| JPWO2011019072A1 (en) * | 2009-08-12 | 2013-01-17 | ヒューマン・メタボローム・テクノロジーズ株式会社 | Biomarker for depression, method for measuring biomarker for depression, computer program, and storage medium |
| AU2010324544A1 (en) * | 2009-11-27 | 2012-06-21 | Baker Idi Heart And Diabetes Institute Holdings Limited | Lipid biomarkers for stable and unstable heart disease |
| EP2348318A1 (en) * | 2010-01-21 | 2011-07-27 | Equipo Ivi Investigación, S.L. | Diagnostic method for endometrial receptivity |
| EP2385374B2 (en) * | 2010-05-05 | 2018-02-28 | Zora Biosciences OY | Lipidomic biomarkers for atherosclerosis and cardiovascular disease |
| CN103054843B (en) * | 2012-11-23 | 2015-04-22 | 华北制药集团新药研究开发有限责任公司 | Application of altenusin compound and pharmaceutically acceptable salt thereof in preparation of medicaments for treating FXR-mediated diseases |
| CN104297355B (en) * | 2013-07-17 | 2017-02-08 | 中国科学院大连化学物理研究所 | Simulative-target metabonomics analytic method based on combination of liquid chromatography and mass spectrum |
| US20150090010A1 (en) * | 2013-09-27 | 2015-04-02 | Chang Gung University | Method for diagnosing heart failure |
| CN104614291B (en) * | 2015-01-13 | 2018-01-16 | 江苏普瑞姆纳米科技有限公司 | Assess mark and the method that airborne fine particulate matter exposure is acted on organismal toxicity |
-
2016
- 2016-01-25 CN CN201610048916.3A patent/CN105445408B/en active Active
- 2016-01-28 WO PCT/CN2016/072440 patent/WO2017128162A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150376704A1 (en) * | 2009-12-09 | 2015-12-31 | Cleveland Heartlab, Inc. | Biomarker assay for diagnosis and classification of cardiovascular disease |
| CN103154742A (en) * | 2010-06-20 | 2013-06-12 | 佐拉生物科学公司 | Lipidomic biomarkers for identification of high-risk coronary artery disease patients |
| CN102901789A (en) * | 2012-09-21 | 2013-01-30 | 中国药科大学 | Determination method of serum metabolic marker for early diagnosis of diabetic nephropathy. |
| JP2015065979A (en) * | 2013-09-26 | 2015-04-13 | テルモ株式会社 | Guide wire |
| US20150272999A1 (en) * | 2014-03-31 | 2015-10-01 | Banner Health | Retrograde coronary vein delivery of stem cells |
| KR20150059644A (en) * | 2015-05-01 | 2015-06-01 | 학교법인 동아학숙 | composition for diagnosing coronary artery disease comprising CD31 or fragment thereof |
Non-Patent Citations (1)
| Title |
|---|
| 魏晓真等: "不稳定型心绞痛 急性心肌梗死患者血清磷脂变化的临床意义", 《中国急救医学》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108072729A (en) * | 2016-11-18 | 2018-05-25 | 华北制药集团新药研究开发有限责任公司 | A kind of method for measuring shellfish cholic acid piece dissolution rate difficult to understand |
| CN106546721A (en) * | 2016-11-25 | 2017-03-29 | 南京拓睿谱基因科技有限公司 | Glandular cystitiss and Diagnosis of Bladder diacritics thing, diagnostic reagent or test kit |
| CN107677825A (en) * | 2017-10-06 | 2018-02-09 | 南京普氟生物检测技术有限公司 | A kind of purposes for the tumor diagnosis composition of diagnosis of cervical cancer and for preparing diagnostic kit |
| CN107677825B (en) * | 2017-10-06 | 2019-04-23 | 汉氏联合（天津）干细胞研究院有限公司 | A kind of tumor diagnosis composition for diagnosis of cervical cancer and the purposes for being used to prepare diagnostic kit |
| CN107589194A (en) * | 2017-11-02 | 2018-01-16 | 南京拓睿谱基因科技有限公司 | A kind of lung cancer early diagnosis small molecule mark and the application in terms of diagnosis |
| CN107589194B (en) * | 2017-11-02 | 2019-01-29 | 武汉迈特维尔生物科技有限公司 | A kind of lung cancer early diagnosis small molecule marker and the application in terms of diagnosis |
| CN111505131A (en) * | 2020-01-02 | 2020-08-07 | 东莞东华医院有限公司 | Clinical model for predicting coronary heart disease plaque instability based on serum metabonomics change |
| CN112305123A (en) * | 2020-10-30 | 2021-02-02 | 河北医科大学第二医院 | Application of small molecular substance in atherosclerotic cerebral infarction |
| CN118258931A (en) * | 2024-05-29 | 2024-06-28 | 山东第一医科大学附属省立医院(山东省立医院) | Metabolic marker combination for early warning female atherosclerosis, application thereof and early warning method |
| CN118258931B (en) * | 2024-05-29 | 2024-08-16 | 山东第一医科大学附属省立医院(山东省立医院) | Metabolic marker combination and detection kit for early warning female atherosclerosis |
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| WO2017128162A1 (en) | 2017-08-03 |
| CN105445408B (en) | 2018-06-12 |
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