Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip

A nanoparticle and immunochromatographic technology, applied in biological testing, fluorescence/phosphorescence, measurement devices, etc., can solve the problems of difficult recording and storage of test results, low detection sensitivity, and difficult quantitative detection, and achieve easy operation and high sensitivity. , the effect of high sensitivity

Inactive Publication Date: 2005-07-27
XIAMEN UNIV
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  • Summary
  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of colloidal gold technology is that the detection sensitivity is low, it is difficult to carry out quantitative detection, and visual detection makes it difficult to record and save the detection results

Method used

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  • Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip
  • Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip
  • Immune chromatography with fluorescent rare earth nanometer particle as marker and detecting testing paper strip

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: Preparation of HBsAg Rapid Detection Test Strips

[0045] In this embodiment, HBsAg is used as the detection object, and the bonded fluorescent rare earth complex nanoparticles with a diameter of about 50nm and nitrocellulose chromatography membrane with amino groups on the surface are taken as examples. Through labeling, coating and assembly of various components, The finished product of the immunochromatographic detection test strip using fluorescent rare earth complex nanoparticles as markers is prepared.

[0046] (1) Labeling of anti-HBsAg monoclonal antibody

[0047] Method 1: Directional Markup

[0048] Take 500 μL (3.6 mg / mL) of affinity-pure anti-HBsAg monoclonal antibody, dialyze against 0.01 M sodium acetate buffer (pH5.2) at 4 °C for 6 h; then add NaIO 4 , to oxidize the antibody. After 20 minutes, dialyze against 0.01M pH5.2 sodium acetate buffer again; then add 500 μL (18 mg / mL) of BHHCT-Eu particles with amino groups on the surface, mix well,...

Embodiment 2

[0062] Example 2: Detection of biotin by biotin-nucleophile system simulation competition method

[0063] This example combines streptavidin mimetic capture antibody with biotinylated Tb 3+ The complex nanoparticles were used as markers, and the biotin was simulated by the competition method. The particle diameter used in the example is about 50nm.

[0064] (1)Tb 3+ Biotinylation of Complex Nanoparticles

[0065] method one:

[0066]Take 500 μL of BPTA-Tb particles with amino groups on the surface to be labeled (suspended in absolute ethanol), add a little biotin-mPEG-SPA, dissolve, mix well, and react at room temperature for 3 h; then, add absolute ethanol and 50 mM tris 7.8 The particles were washed 3 times with buffer, and then washed with 500 μL 50 mM tris 7.8 (containing 0.9% NaCl, 0.2% BSA, 0.1% NaN 3 ) for suspension.

[0067] Method Two:

[0068] Take 500 μL of BPTA-Tb particles with amino groups on the surface to be labeled (suspended in ultrapure water), treat...

Embodiment 3

[0077] Embodiment 3: One-step multi-inspection detection

[0078] The one-step multi-detection project was simulated with the HBsAg detection system and the biotin-nucleophile system. The HBsAg detection system of the first detection zone uses a pair of monoclonal antibodies at different sites as the capture antibody and the labeled antibody, in which the labeled antibody and Eu 3+ The complex nanoparticles are cross-linked and detected by the sandwich method. The biotin-nucleophile system of the second detection zone uses nucleophilic analog capture antibody to biotinylated Tb 3+ The complex nanoparticles were used as markers, and the biotin was simulated by the competition method. Rabbit anti-mouse IgG was used as the detection band to capture the antibody. The marker pad also contains anti-HBsAg monoclonal antibody labeled Eu 3+ Complex nanoparticles and biotin-labeled Tb 3+ complex nanoparticles.

[0079] (1) Labeling of anti-HBsAg monoclonal antibody

[0080] Same ...

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Abstract

An immune chromatographic method includes fixing capture antigen or antibody directly on chromatographic film (PH) by physical absorption means, adding sample liquid and label antibody on sample cushion side of PF and forming immmune composite through immune reaction, detecting formed detection and control belts, sticking PE, sample loading cushion, label cushion and water absorption cushion in sequence on backing. The test paper is also disclosed.

Description

technical field [0001] The invention relates to an immunochromatographic detection method, in particular to a rapid immunochromatographic detection method using fluorescent rare earth complex nanoparticles as markers and a detection test strip thereof. The fluorescent rare earth complex nanoparticles are silicon nanoparticles doped or bonded with fluorescent rare earth complexes. Background technique [0002] Immunoassays are versatile methods for the detection of antibodies, antigens, and haptens. There are many detection modes of immunoassay, among which solid-phase immunoassay and immunochromatography are routine clinical methods. The solid-phase immunoassay method includes sample incubation and washing steps. This method has the advantages of high sensitivity and can be used for semi-quantitative or even quantitative detection. In particular, the detection mode of the 96-microwell plate is convenient for the detection of a large number of samples. It is the current clin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/52G01N33/533G01N33/543G01N33/547
Inventor 李庆阁许晔
Owner XIAMEN UNIV
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