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Immune chromatography fluorescence reagent strip for detecting cardiac troponin and preparation method thereof

A technique of cardiac troponin and immunochromatography, which is applied in the field of clinical medical testing to achieve the effects of saving costs, saving materials, and shortening detection time

Inactive Publication Date: 2014-12-24
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no in vitro rapid diagnostic product guided by the principle of metal-enhanced immunochromatographic fluorescence on the market

Method used

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  • Immune chromatography fluorescence reagent strip for detecting cardiac troponin and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 Preparation of tracer particle binding pad

[0025] Spray the tracer particle-labeled antibody / antigen marker evenly on the processed sample pad, and dry it at 25°C for 4 hours. The preparation of the tracer particle can be divided into the preparation of antibody IgG-fluorescein and the preparation of metal particle-antibody - Two-step preparation of the fluorescent complex:

[0026] A. Preparation of antibody IgG-fluorescein: Dissolve antibody IgG in 20-100Mmol PBS buffer, dissolve activated fluorescein (FITC, Cy5, etc.) in dimethyl sulfoxide (DMF), then mix antibody IgG with fluorescein Mix at 1:1~1:3, fully stir and react for 1 hour, then place the solution in a dialysis bag for gradient fluid dialysis for 3 times, each time for 6~8 hours, and the obtained solution is the antibody IgG-fluorescein complex;

[0027] B. Preparation of metal particle-antibody-fluorescent complex: Mix 50~200nm nanometer metal particles and antibody IgG-fluorescein at a ra...

Embodiment 2

[0028] Embodiment 2 Preparation of an immunochromatographic fluorescent reagent strip for detecting cardiac troponin

[0029] An immunochromatographic fluorescent reagent strip for detecting cardiac troponin, comprising a bottom plate 1, on which a sample pad 2, a tracer particle binding pad 3, a nitrocellulose membrane 4 and a water-absorbing pad 5 are sequentially pasted; Points 2-5 need to overlap by 1~2mm to ensure that the test sample passes through the test area and reaches the absorbent pad smoothly. The preparation method of each component is as follows:

[0030] 1) Preparation of sample pad: The sample pad can be made of glass fiber or polyester, and 5mg / ml of protein (BSA, casein, etc.) can be dissolved in buffer (PBS, Tris or glycine, etc.), and then add a small amount of surface activity agent (Tween20, etc.), adjust the pH to 6-8, soak the glass fiber or polyester material for 2-4 hours, take it out and dry it at 25°C for 8 hours;

[0031] 2) Preparation of trace...

Embodiment 3

[0037] Example 3 Detection of cardiac troponin protein using the reagent described in this utility

[0038] In embodiment 2 such as figure 1 The shown reagent strips are cut into 5mm widths and installed on a suitable reagent strip support shell, and the sample pad 2 faces the sample injection hole, and presses the shell tightly. Then the sample detection operation steps are as follows: (1) collect serum, plasma or whole blood samples and return to room temperature; (2) drop 100ul of the sample to be tested onto the sample pad 2 for reaction; (3) read the results, and after 90s The above reagent strips are placed in the immunoquantitative analyzer to read the results of the corresponding samples.

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Abstract

The invention discloses an immune chromatography fluorescence reagent strip for detecting cardiac troponin and a preparation method thereof, belonging to the field of clinical medicine detection. The reagent strip comprises a baseboard, a sample mat, a tracer particle combining mat, a nitrocellulose membrane and a water absorption mat; the sample mat, the tracer particle combining mat, the nitrocellulose membrane and the water absorption mat are orderly connected and fixed on the baseboard in the horizontal direction. The tracer particle combining mat is provided with the cTnI monoclonal antibody-fluorescence particle compound adsorbed layer covered on the surface of the nanocrystalline metal particle; the nitrocellulose membrane is provided with the detection line composed of the cTnI monoclonal antibody or polyclonal antibody capture molecule and the quality control line composed of the rabbit antimouse IgG antibody. The immune chromatography fluorescence reagent strip for detecting cardiac troponin is short in detection time, high in specificity and sensitivity and more accurate in detection result by improving the fluorescence intensity through the nanocrystalline metal particle.

Description

technical field [0001] The invention relates to the field of clinical medical examination, in particular to an immunochromatographic fluorescent reagent strip for detecting cardiac troponin and a preparation method thereof. Background technique [0002] Immunochromatography is a method of in vitro diagnostic reagents, which has the characteristics of low price, convenient detection and rapid detection. It combines technical methods such as affinity technology, labeling technology, imprinting technology and chromatography technology. [0003] Currently used immunochromatographic reagent methodology can be divided into colloidal gold method and fluorescence method. Fluorescence techniques are gradually becoming dominant in immunochromatography. However, this technology has disadvantages such as low sensitivity, long detection time, and high antibody cost. It is necessary to further improve the detection sensitivity, reduce the detection time, and save the amount of antibody ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6887
Inventor 金晶杜腾飞黄力苏恩本王勇陈伟
Owner GETEIN BIOTECH
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