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Metabolic marker combination for evaluating risks of cardiovascular disease of subject and application thereof

A technology for metabolic markers, cardiovascular and cerebrovascular diseases, applied in instruments, measuring devices, scientific instruments, etc., can solve problems such as difficult to meet the medical industry

Active Publication Date: 2021-03-16
HUMAN METABOLOMICS INST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At the same time, the current traditional biomarker detection methods include thin-layer chromatography, liquid chromatography, immunochemical methods, and gas chromatography. High standards of coverage and dynamic range
Existing research and detection methods for cardiovascular and cerebrovascular diseases based on mass spectrometry mainly use non-targeted relative quantitative or semi-quantitative methods, or absolute quantitative methods of a single type of compound for disease diagnosis or evaluation detection, resulting in the accuracy of the detection results and Comprehensiveness has certain limitations, and there is still a certain distance from truly effective clinical mass spectrometry detection and risk assessment

Method used

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  • Metabolic marker combination for evaluating risks of cardiovascular disease of subject and application thereof
  • Metabolic marker combination for evaluating risks of cardiovascular disease of subject and application thereof
  • Metabolic marker combination for evaluating risks of cardiovascular disease of subject and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, blood sample collection

[0062] Vacuum blood collection tubes with clinical EDTA anticoagulation were used to collect 3-5ml fasting venous blood intravenously from patients with coronary heart disease and healthy controls. Plasma was separated at 1500 g x 15 minutes within 1 hour after collection. Aliquot (150 microliters) into 200 microliter centrifuge tubes, quickly store at -80°C for future use and complete information registration. These include: (1) training set: 183 clinical plasma samples of patients with coronary heart disease and 94 samples of healthy control plasma; (2) validation set: 147 clinical plasma samples of coronary heart disease patients and 94 control plasma samples of healthy subjects.

Embodiment 2

[0063] Example 2. Preparation of Calibrator (Standard) Curve Working Solution, Internal Calibrator and Quality Control

[0064] The standard substance of the marker to be tested was dissolved into a 0.1 mM stock solution in a solvent of isopropanol: acetonitrile ratio of 9:1. Then further dilute with 50 mg / mL bovine serum albumin (BSA, Aladdin) solution to form a mixed calibrator (standard) curve working solution.

[0065] Exemplarily, ceramide Cer d18:1 / 16:0, ceramide Cerd18:1 / 18:0, ceramide GlcCer d18:1 / 12:0 and sphingosine-1-phosphate can be added to the mixed calibrator working solution Alcohol, the concentration points are 2μM, 1μM, 0.4μM, 0.2μM, 0.08μM and 0.04μM; ceramide Cer d18:1 / 24:0, ceramide Cerd18:1 / 24:1, trihexosylceramide d18:1 / 24:), and phenylacetylglutamine at individual concentration points of 10 μM, 5 μM, 2 μM, 1 μM, 0.4 μM μM, and 0.2 μM; also works in mixed calibrator Trimethylamine oxide, trimethylamine, choline, L-carnitine, betaine, and creatinine we...

Embodiment 3

[0068] Example 3. Pretreatment of plasma samples and extraction of diagnostic marker compositions

[0069] Take 10 microliters of the above-mentioned 518 cases of plasma samples, calibrators (standards) curve working solution and quality control products respectively into the V-bottom 96-well plate, then add 190 microliters of the internal calibrator working solution, and affix the aluminum seal Membrane, shake and mix at 650 rpm for 20 minutes. Then centrifuge at 4000×g for 20 minutes, and take 100 microliters of supernatant to be detected by high performance liquid chromatography-mass spectrometry.

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Abstract

The invention discloses a metabolic marker combination for evaluating the risks of cardiovascular and cerebrovascular disease of a subject. The metabolic marker combination comprises ceramide Cer d18:1 / 16:0, ceramide Cer d18:1 / 18:0, ceramide Cer d18:1 / 24:0, phenylacetyl glutamine, trimethylamine, glycine betaine and choline. The cardiovascular and cerebrovascular diseases are selected from coronary heart disease, atherosclerosis, atrial fibrillation and heart failure. The metabolic marker combination disclosed by the invention has the advantages of high sensitivity, good specificity, quantification, high detection flux and the like in the aspect of evaluating or predicting the risks of cardiovascular and cerebrovascular diseases.

Description

technical field [0001] The present invention relates to the field of cardiovascular and cerebrovascular diseases, in particular to a metabolic marker for assessing the risk of cardiovascular and cerebrovascular diseases of subjects and its application. Background technique [0002] Cardiovascular and cerebrovascular diseases are collectively referred to as cardiovascular and cerebrovascular diseases, and generally refer to ischemic or hemorrhagic diseases of the heart, brain, and systemic tissues caused by hyperlipidemia, blood viscosity, atherosclerosis, and high blood pressure. Diseases, including hypertension (increased blood pressure), coronary artery disease (heart attack), cerebrovascular disease (stroke), peripheral vascular disease, heart failure, rheumatic heart disease, congenital heart disease, and cardiomyopathy, among others. [0003] The morbidity and mortality of cardiovascular and cerebrovascular diseases are high, and the pathogenesis is complex. Most of th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8813G01N2030/884
Inventor 贾伟谢国祥林志龙
Owner HUMAN METABOLOMICS INST INC
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