Kit for evaluating coronary artery disease

A subject, ceramide technology, applied in the field of kits, can solve problems such as hindering the development of clinically effective immunoassay methods, and achieve the effects of good sensitivity and specificity, high throughput and good selectivity

Pending Publication Date: 2022-02-18
江苏豪思生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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 Lipid Res48, 968-975, doi: 10.1194 / jlr.D600043-JLR200(2007).)

Method used

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  • Kit for evaluating coronary artery disease
  • Kit for evaluating coronary artery disease
  • Kit for evaluating coronary artery disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1: Subject sample collection, preservation and transportation

[0093] 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.

[0094] (1) Sample collection

[0095] 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.

[0096] (2)Sample preservation

[0097] 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.

[0098] (3) Sample transportation

[0099] Plasma was shipped at 4°C on blue ice.

Embodiment 2

[0100] Example 2: Pretreatment of the plasma sample collected in Example 1 before LC-MS / MS detection (protein rapid precipitation method)

[0101] 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 can also include: for making Cer(d18:1 / 16:0), Cer(d18:1 / 18:0), Cer(d18:1 / 24:1) and Cer(d18:1 / 24:0) Four kinds of pure ceramides (commercially available products) of the standard curve of four kinds of ceramides or four kinds of pure ceramides storage solution prepared by methanol solvent (the concentration of the storage solution can be 500pmol / μl); Four kinds of ceramide internal standard (IS, i.e. four kinds of ceramide corresponding deuterated substances) solutions prepared by methanol solvent, the concentrations are respectively D 7 -Cer d18:1 / 16:0: 0.125 pmol / μl, D 7 -Cer d18:1 / 18:0 0.05 pmol / μl, D ...

Embodiment 3

[0103] Example 3: Optimization of LC-MS / MS detection and analysis conditions.

[0104] 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.

[0105] 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.

[0106] Before testing the subjects' plasma samples, the testing conditions were comprehensively optimized to obtain Cerd18:1 / 16:0, D 7 -Cer d18:1 / 16:0, Cer d18:1 / 18:0, D 7 -Cer d18:1 / 18:0, Cer d18:1 / 24:0, D 7...

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Abstract

The invention provides a kit for evaluating coronary artery diseases. The kit comprises a detection reagent and a specification, wherein the detection reagent is used for detecting the concentrations of Cer (d18:1 / 16:0), Cer (d18:1 / 18:0), Cer (d18:1 / 24:0) and Cer (d18:1 / 24:1) in a sample of a tested object. 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

[0001] This application is a divisional application of the invention patent application with application number 201711338605.1, and the application date is December 14, 2017. technical field [0002] The invention relates to a kit, in particular to a kit for evaluating coronary artery disease. Background technique [0003] Cardiovascular disease has become the disease with the highest human mortality rate in the world, and the annual death rate of cardiovascular disease has surpassed other diseases, becoming the first cause of death. In 2015, about 177 million people died from cardiovascular disease, accounting for 31% of the global death toll, of which 74 million people died from coronary artery disease. With the development of my country's economic level, the improvement of people's living standards, the change of diet structure and the rapid aging of the population, the morbidity and mortality of cardiovascular diseases are on the rise. The 2016 "China Cardiovascular Dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/72G01N30/06
CPCG01N30/72G01N30/06
Inventor 栗琳崔雅轩崔文宏张丽丽郝敬雨姜平
Owner 江苏豪思生物科技有限公司
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