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Fluorescence immune chromatography test paper and preparing method and application thereof

A fluorescence immunochromatography and test paper technology, applied in the field of medical testing, can solve the problems of low detection sensitivity, inability to use quantification, fluorescence quenching, etc., and achieve the effect of high sensitivity and good accuracy

Inactive Publication Date: 2009-09-09
沈鹤柏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to provide a fluorescent immunochromatographic detection test paper to overcome that the UCP particles used in the prior art are not packaged, which is prone to fluorescence quenching, and the immunological detection using UCP particles is a qualitative detection, which cannot Good enough for quantification and detects defects with low sensitivity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of nucleocapsid FITC-alpha-fetoprotein (AFP) antibody complex

[0038] Surface siliconization of fluorescein particles: In the ethanol system, add 0.1-10mg fluorescein isothiocyanate FITC and 0.1-10μL N-(2-aminoethyl)-3-aminopropyltrimethoxysilane, 15- Stir at 50°C for 3-10 hours, then add 0.1-10mL water, 0.1-10mL 15%wt ammonia water, 0.1-10mL tetraethyl orthosilicate (TEOS), and continue to react for 3-10 hours. Prime particles.

[0039] Surface amination of fluorescein: Take the fluorescein particles with siliconized surface above, add 10-50mL of a mixed solution consisting of 1:3-3:1 (V / V) methanol and glycerin, ultrasonically disperse, and then add 10-500μL N- (2-aminoethyl)-3-aminopropyltrimethoxysilane, stirred at a constant temperature of 15-50°C for 5-10 hours, washed with ethanol, and dried to obtain surface aminated fluorescein particles.

[0040] Conjugation to AFP antibody using surface aminated core-shell FITC:

[0041] 1. Add 2 mg...

Embodiment 2

[0046] Example 2: Preparation of double-antibody sandwich AFP antigen detection test paper

[0047] 1. Preparation of the sample pad: select cellulose film as the sample pad material, cut it into strips of 1.5×30.0cm, put it into the sample pad blocking solution (0.03mol / L PB (pH=7.2), containing 5mg / L mLBSA, 0.1% Triton X-100) for 30min, and dried at 37°C for later use.

[0048] 2. Preparation of conjugation pads: select glass cellulose membrane as the conjugation pad material, cut it into 1.0×30.0 cm strips, centrifuge the core-shell type FITC-AFP antibody complex prepared in Example 1, and settle Add 0.03mol / LPB (pH=7.2), containing 1% sucrose, 1% BSA) to the mixture, mix thoroughly, add to the strip, and dry at 37°C.

[0049] 3. Preparation of NC: Cut the NC membrane into 2.5×30.0cm strips, spray AFP monoclonal antibody and goat anti-mouse IgG antibody on different positions on the NC membrane with a sampler, as the detection strip and quality control belt, and dry at 37...

Embodiment 3

[0051] Example 3: Tetramethylrhodamine TRITC and tetraethylrhodamine RB200 were used instead of FITC in Example 1-2 to prepare double-antibody sandwich AFP antigen detection test paper.

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Abstract

The invention relates to fluorescence immune chromatography test paper which is formed by mutually and sequentially overlapping a sample pad, a combination pad, an antibody carrying film and water absorbing paper on a lining board with adhesive, wherein the combination pad is coated with a fluorescence material antibody 1 composite, a fluorescence-marked material is fluorescein granules or fluorescence material converted by rare earth, the antibody carrying film is a cellulose nitrate film or a nylon film and is respectively connected with an antibody 2 and the T line and the C line of a secondary antibody, the fluorescence material is fluorescein granules or the fluorescence granules converted by the rare earth, and the fluorescein granules are selected from isosulfocyanic acid fluorescein or tetramethyl isosulfocyanic acid rhodamine or tetraethyl rhodamine, and the surface of the fluorescence-marked material is aminated and combined with the antibody by cross-linking reaction. The fluorescence quantitative measuring system is used for detecting the fluorescence strength of the areas of the T line and the C line, and the quantitative detection is completed by the standard curve of the antigen concentration. The invention is suitable for the immune detection of tumor marked objects of urinal fiber-connected protein, and the like.

Description

technical field [0001] The invention belongs to the field of medical testing, in particular to a fluorescent immunochromatographic detection test paper. Background technique [0002] Tumor is a disease with high morbidity and high mortality that threatens human health. A large number of research and prevention data have confirmed that early diagnosis and early treatment are the most effective ways to prevent and treat tumors and reduce mortality. For this reason, people have conceived a variety of measures and approaches. For patients with early asymptomatic tumors, tumor markers are often used as one of the methods for early diagnosis of tumors. Therefore, the constant search for new tumor markers and their detection methods has become a research hotspot in the academic circles. [0003] At present, clinically, tumor markers are mainly detected by immunoassay techniques, such as enzyme-linked immunoassay (ELISA), radioimmunoassay (RIA), chemiluminescence assay (CLIA), etc....

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/52G01N33/533
Inventor 沈鹤柏赵露晶马经纬
Owner 沈鹤柏
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