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Kit for evaluating coronary artery disease and application thereof

A coronary artery disease and kit technology, applied in the field of kits, can solve the problems of lack of selectivity, low precision and accuracy, low throughput, etc., and achieve good sensitivity and specificity, high precision and accuracy, selective effect

Pending Publication Date: 2022-03-04
湖南豪思生物科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the methods are designed for research work rather than clinical use, and the common disadvantages of these methods are that they cannot analyze ceramides at the molecular species level, and lack selectivity, low throughput, precision and The accuracy is low, so it cannot meet the requirements of clinical laboratories
It is also impossible to use traditional immunoassay methods to specifically detect ceramide and phosphatidylcholine compounds, only to recognize a class of ceramide molecules, which only differ in the length of the fatty acid chain
Therefore, the low selectivity of existing antibodies hinders the development of clinically effective immunoassay methods (Krishnamurthy, K., Dasgupta, S. & Bieberich, E. Development and characterization of a novel anti-ceramide antibody. J LipidRes 48, 968-975, doi :10.1194 / jlr.D600043-JLR200(2007).)

Method used

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  • Kit for evaluating coronary artery disease and application thereof
  • Kit for evaluating coronary artery disease and application thereof
  • Kit for evaluating coronary artery disease and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] Example 1: Subject sample collection, preservation and transportation

[0110] Reagents and test supplies for collecting subjects' samples can be included in the kit of the present invention, such as the EDTA anticoagulant tube (purple cap) used below.

[0111] (1) Sample collection

[0112] Collect 3ml of the subject's blood through venous blood collection, and separate the plasma by centrifugation (centrifugation conditions: 1300g, 10min) within 6 hours after blood collection; preferably, EDTA anticoagulant tubes (purple cap) are the first choice to separate plasma.

[0113] (2)Sample preservation

[0114] If it cannot be used for detection immediately, the separated plasma must be stored at 2-8°C or -20°C and protected from light.

[0115] (3) Sample transportation

[0116] Plasma was shipped at 4°C on blue ice.

Embodiment 2

[0117] Example 2: Pretreatment of the plasma sample collected in Example 1 before LC-MS / MS detection (protein rapid precipitation method)

[0118] Reagents and test supplies for pretreatment of plasma samples can be included in the kit of the present invention, such as: protein precipitation (PPT) solvent: ethyl acetate: isopropanol (2:8, v / v), this The kit of the invention may also include: for making Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 20:0), Cer(d18:1 / 22:0), Cer(d18:1 / 24:0), Cer(d18:1 / 24:1) and PC aa C36:6, PC aa C38:0, PC aa C38:6) standard curve of phospholipids The pure product of the compound (commercially available product) or the pure product storage solution prepared by methanol solvent (the concentration of the storage solution can be 500pmol / μl); the four ceramide internal standards (IS, that is, the four ceramides corresponding to the four ceramides) prepared by methanol solvent Deuterium) solution, its concentration is D 7 -Cer d18:1 / 16:0: 0.125 pmol / μ...

Embodiment 3

[0120] Example 3: Optimization of LC-MS / MS detection and analysis conditions.

[0121] The optimized LC-MS / MS detection and analysis conditions and equipment used in this example can be described in the instructions of the kit of this application. Or / and the reagents needed to optimize the detection and analysis conditions of LC-MS / MS are used as a part of the reagents in the kit of the present invention.

[0122] LC-MS / MS analysis was performed on a Thermo SCIENTIFIC TSQ Quantitative mass spectrometer coupled with a Thermo SCIENTIFIC UPLC Ultimate3000. Electrospray ionization (ESI) in positive ion mode employs multiple reaction monitoring (MRM). Instrument control and data acquisition were controlled using the Thermo SCIENTIFIC TSQ Quantitative companion software.

[0123] Before the detection work on the subject's plasma samples, the detection conditions were comprehensively optimized to obtain D 7 -Cer d18:1 / 16:0, D 7 -Cer d18:1 / 16:0, D 7 -Cer d18:1 / 18:0, D 7 -Cer d18...

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Abstract

The invention provides a kit for evaluating coronary artery diseases. The kit comprises detection reagents for detecting concentrations of ceramide compounds Cer (d18: 1 / 16: 0), Cer (d18: 1 / 18: 0), Cer (d18: 1 / 20: 0), Cer (d18: 1 / 22: 0), Cer (d18: 1 / 24: 0) and Cer (d18: 1 / 24: 1) in a sample of a tested object and phosphatidylcholine compounds PC aa C36: 6, PC aa C38: 0 and PC aa C38: 6. The kit disclosed by the invention can be efficiently and accurately used for coronary artery disease diagnosis, risk stratification, illness state monitoring and curative effect evaluation, is relatively good in sensitivity and specificity, and can meet the requirements of clinical diagnosis and treatment of coronary artery diseases.

Description

technical field [0001] The invention relates to a kit, in particular to a kit for evaluating coronary artery disease and its application. Background technique [0002] Given the high morbidity and mortality associated with coronary artery disease, the prevention of fatal and nonfatal myocardial infarction in patients with coronary artery disease remains a clinical challenge. The annual mortality rate in patients with stable coronary artery disease is between 1-3%, and the non-fatal morbidity rate is between 1-2%. Among patients with acute coronary syndrome who survived the acute condition, the mortality rate from myocardial infarction was significantly increased (especially within the first year). However, morbidity risk varies greatly among individual patients, requiring effective tools for diagnosis, risk stratification, disease monitoring, and efficacy evaluation to improve patient identification and management. [0003] Coronary artery disease markers are biologically ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N30/72G01N30/02
CPCG01N33/50G01N30/02G01N30/72
Inventor 栗琳张丽丽肖冰心王春静丁亮周立
Owner 湖南豪思生物科技有限公司
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