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Heart-type fatty acid binding protein content detection kit and preparation method thereof

A fatty acid binding and detection kit technology, which is applied in the measurement of color/spectral properties, etc., can solve the problems of long time consumption, cumbersome operation and low sensitivity of enzyme-linked immunosorbent assay, and is conducive to large-scale promotion and application, detection operation. Simple and highly accurate results

Active Publication Date: 2015-04-15
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, H-FABP detection methods include enzyme-linked immunoassay (ELISA), radioimmunoassay (RIA), immunochromatography, etc., but these detection methods have some defects, which limit their large-scale application in clinical diagnosis , such as: enzyme-linked immunoassay is time-consuming, cumbersome to operate, and poor in repeatability; radioimmunoassay requires the use of specific instruments and is contaminated by radioactive substances; immunochromatography is mostly used for qualitative detection, and the method has low sensitivity
Latex-enhanced immune turbidimetry has the advantages of simple operation, rapidity, high sensitivity, and can be applied to automatic biochemical analyzers. Although there are some H-FABP kits on the market based on latex-enhanced immune turbidimetry, these reagents or There are more or less problems such as insufficient detection linear range, insufficient detection sensitivity, poor reagent storage stability, and poor anti-interference ability during the detection process.

Method used

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  • Heart-type fatty acid binding protein content detection kit and preparation method thereof
  • Heart-type fatty acid binding protein content detection kit and preparation method thereof
  • Heart-type fatty acid binding protein content detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Preliminary Example 1 Preparation of Latex Particles Coated with Heart-Type Fatty Acid Binding Protein Antibody

[0042] (1) Activated latex: Take 1 mL of polystyrene latex solution (purchased from PolyMicrospheres) with a concentration of 10% and a particle size of 90 nm in a 50 mL centrifuge tube, then add 9 mL of 0.12M phosphate buffer solution to the centrifuge tube, and mix well , to obtain a diluted latex solution; accurately weigh 50 mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and dissolve it in 1 mL of 0.12M phosphate buffer solution, and add this solution to complete the dilution In the latex solution, oscillate and mix evenly, and place on a 25°C constant temperature shaker, 200 rpm, and react for 1h; On a shaker, 200 rpm, react for 10 minutes;

[0043] (2) Sensitized latex: Add 2 mL of 1 mg / mL heart-type fatty acid binding protein polyclonal antibody (purchased from Wuhan Huamei Bioengineering Co., Ltd.) to the solution completed in step (1), ...

Embodiment 2

[0046] Preliminary Example 2 Preparation of latex particles coated with heart-type fatty acid binding protein antibody

[0047] (1) Activated latex: Take 1 mL of polystyrene latex solution with a concentration of 10% and a particle size of 200 nm (purchased from PolyMicrospheres) in a 50 mL centrifuge tube, then add 9 mL of 0.12M phosphate buffer solution to the centrifuge tube, and mix well , to dilute the latex solution. Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and dissolve it in 1mL of 0.12M phosphate buffer solution, and add this solution into the diluted latex solution, shake Mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm, and react for 1 hour; after the reaction is completed, add 2 μL of 2-mercaptoethanol to the above system, shake and mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm min, reaction 10min;

[0048] (2) Sensitized latex: Add 2 mL of 1 mg / mL heart-type fatty acid binding pr...

Embodiment 3

[0051] Preliminary Example 3 Preparation of Latex Particles Coated with Heart-type Fatty Acid Binding Protein Antibody

[0052] (1) Activated latex: Take 1 mL of polystyrene latex solution with a concentration of 10% and a particle size of 120 nm (purchased from PolyMicrospheres) in a 50 mL centrifuge tube, then add 9 mL of 0.12M phosphate buffer solution into the centrifuge tube, and mix well , to dilute the latex solution. Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and dissolve it in 1mL of 0.12M phosphate buffer solution, and add this solution into the diluted latex solution, shake Mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm, and react for 1 hour; after the reaction is completed, add 2 μL of 2-mercaptoethanol to the above system, shake and mix evenly, and place on a constant temperature shaker at 25°C, 200 rpm min, reaction 10min;

[0053] (2) Sensitized latex: Add 2 mL of 1 mg / mL heart-type fatty acid binding ...

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Abstract

The invention discloses a heart-type fatty acid binding protein content detection kit and a preparation method thereof. The kit disclosed by the invention is formed by a reagent I and a reagent II which are mutually independent; the reagent I comprises the following components of: a biological buffering agent, a surfactant, a coagulation accelerator, a preservative, a stabilizing agent, a blocking agent, a chelating agent and water; the reagent II comprises the following components of: latex grains covered by a heart-type fatty acid binding protein antibody, a biological buffering agent, a chelating agent, a surfactant, a preservative, a suspending aid, a sealing agent, a stabilizing agent and water; and the grain diameters of the latex grains covered by the heart-type fatty acid binding protein antibody are within 90-200nm. The detection kit disclosed by the invention has a series of advantages of good stability, strong specificity, high sensitivity, strong anti-interference capability on rheumatoid factors, good linear range, simplicity and convenience for detection and operation and the like, and is good for large-scale popularization and application.

Description

technical field [0001] The present invention relates to a detection kit for the content of cardiac markers, in particular to a detection kit for the content of heart-shaped fatty acid binding protein and a preparation method thereof. The method for using the protein content belongs to the detection field of the heart-type fatty acid binding protein content. Background technique [0002] Acute coronary syndrome (ACS) is one of the most important acute events in coronary heart disease. It is mainly divided into acute myocardial infarction (AMI), unstable angina (UA) and sudden cardiac death (SCD). Both are higher. However, if high-risk patients can be identified early after onset and reperfusion therapy can be performed, the mortality and prognosis will be significantly improved. Among them, cardiac markers play a crucial role in the diagnosis and prognosis of ACS. [0003] In recent years, research on markers of acute myocardial infarction (AMI) has made great progress. T...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
Inventor 华权高沈鹤霄许可黄爱舒芹鄢宝常向博
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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