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Fluorescence immunochromatographic method for quantitative determination of cardiac troponin I and heart-type fatty acid binding protein

A technology of cardiac troponin and fluorescent immunochromatography, which is applied in biological testing, material inspection products, etc., and can solve problems such as false positive increase, soluble complex error, and blood lipid concentration influence

Inactive Publication Date: 2017-12-19
CHANGZHOU BIOWIN BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this method is that when there is a large excess of antibodies or antigens, soluble complexes will appear to cause errors, and it is easily affected by the concentration of blood lipids. Small particles of lipoproteins can form turbidity, resulting in increased false positives.

Method used

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  • Fluorescence immunochromatographic method for quantitative determination of cardiac troponin I and heart-type fatty acid binding protein
  • Fluorescence immunochromatographic method for quantitative determination of cardiac troponin I and heart-type fatty acid binding protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Preparation of a fluorescent immunoassay kit for cardiac troponin I and heart-shaped fatty acid binding protein

[0029] 1) Coupling of fluorescent dyes and cardiac troponin I antibody

[0030] Mix the fluorescent dye rhodamine with an emission wavelength of 665nm and 1mg / ml of myocardial troponin I monoclonal antibody, react at room temperature for 3h, add 1mol / L hydroxylamine hydrochloride to terminate the reaction, and separate and purify with a chromatographic column or a chromatographic column to obtain fluorescence Dye-modified cardiac troponin I monoclonal antibody, fluorescence emission wavelength is 665nm;

[0031] 2) Coupling of fluorescent dye and heart-shaped fatty acid binding protein antibody

[0032] The fluorescent dye rhodamine with emission wavelength of 665nm was mixed with 1mg / ml cardiotypic fatty acid binding protein monoclonal antibody, reacted at room temperature for 3h, and 1mol / L hydroxylamine hydrochloride was added to terminate the reaction...

Embodiment 2

[0038] Example 2: Detection method of cardiac troponin I and heart-shaped fatty acid binding protein fluorescence immunoassay kit

[0039] 1) Take out the blood sample to be tested;

[0040] 2) Drop 80ul into the sample hole, and react for 15min in forward chromatography;

[0041] 3) Prepare washing buffer solution, pH value is 7.5, buffer system is 20mM phosphate, add bovine serum albumin (concentration is 1%), sucrose (concentration is 10%) and surfactant Triton X-100 ( The concentration is 0.8%), add 50μL to the sample well and let stand for 5min;

[0042] 4) It is placed in a fluorescence quantifier to obtain the fluorescence signal intensity and obtain the concentration of the tested sample.

Embodiment 3

[0043] Example 3: Kit performance evaluation test

[0044] 1) Drawing of standard working curve:

[0045] Take the cardiac troponin I antigen and dilute it with negative serum to a concentration of 60, 50, 30, 20, 10, 5, 1, and 0 ng / mL. Take the heart-shaped fatty acid binding protein antigen and dilute it with negative serum to a concentration of 60, 50, 30, 20, 10, 5, 2, 0 ng / mL specimens, and then determine the troponin I and heart-shaped fatty acid binding protein respectively, and draw the standard working curve with the matching value X and the measured fluorescence value Y. After statistically fitting the standard working curve expression, the regression equation is listed as: Y Cardiac troponin I =0.01219X Cardiac troponin I +0.09901, the square of the fitting coefficient is R 2 =0.982, see the results figure 1 ; Y Heart fatty acid binding protein =0.01213X Heart fatty acid binding protein +0.09628, the square of the fitting coefficient is R 2 =0.982 results see attac...

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Abstract

The invention especially relates to a fluorescence immunochromatographic method for quantitative detection of cardiac troponin I and heart-type fatty acid binding protein, belonging to the technical field of immunodetection and analysis. The fluorescence immunochromatographic method employs blood, an anti-cardiac troponin I antibody, an anti-heart-type fatty acid binding protein antibody, a fluorescence reagent and a detector, wherein the anti-cardiac troponin I antibody is one selected from a group consisting of or a combination of an anti-cardiac troponin I monoclonal antibody and an anti-cardiac troponin I polyclonal antibody; the anti-heart-type fatty acid binding protein antibody is one selected from a group consisting of or a combination of an anti-heart-type fatty acid binding protein monoclonal antibody and an anti-heart-type fatty acid binding protein polyclonal antibody; the fluorescence reagent is a liquid-phase and / or solid-phase material containing a fluorescent substance, and the fluorescent substance is one selected from a group consisting of or a combination of an organic fluorescence dye and a rare earth element fluorescence dye; and the detector is a fluorescence detector. According to the invention, human blood is used as a detection sample, and the method can effectively monitor cardiac troponin I and heart-type fatty acid binding protein in bood, has good specificity, good repeatability and high sensitivity, does not need special instrument and equipment and professional training, and is simple to operate, intelligible and distinct in results, easy to promote and applicable to on-site detection.

Description

Technical field [0001] The invention belongs to the technical field of immune detection and analysis, and particularly relates to a fluorescent immunoassay method for quantitatively detecting cardiac troponin I and heart-type fatty acid binding protein. Background technique [0002] With the improvement of people's quality of life, coronary heart disease has become a major disease affecting people's health. Acute myocardial infarction (AMI) is the main cause of death in coronary heart patients. How to find patients with acute myocardial infarction in time and take thrombolysis and the corresponding interventional treatment as soon as possible is the key to saving patients' lives. Commonly used clinical diagnosis methods of acute myocardial infarction include: clinical manifestations, changes in electrocardiogram and changes in blood biochemical indicators. At present, the specific changes of electrocardiogram is a common method for early diagnosis of acute myocardial infarction...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 李亚星刘凤鸣刘冰
Owner CHANGZHOU BIOWIN BIOPHARM
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