Ultrasensitive and quantitative immunochromatographic device and detection method using same

A technology of immunochromatographic detection and reaction pool, which is applied in the field of biomedicine to achieve the effect of improving detection sensitivity

Inactive Publication Date: 2012-01-04
汤凌霄 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is aimed at the defects of existing immunochromatography technology, to provide a ki...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of Rare Earth Fluorescent Nanoparticle-Antibody Conjugate

[0025] 1) Take 100 μL of rare earth fluorescent nanoparticles (particle size: 300 nm) with a concentration of 10 mg / mL in 900 μL MES (50 mM, Ph6.1), and vortex to mix;

[0026] 2) Add 20mg NHS, 20mg EDC, vortex and mix;

[0027] 3) React at room temperature for 30 minutes;

[0028] 4) Centrifuge at 10000rpm for 10min, discard the supernatant;

[0029] 5) Vortex to resuspend the particles;

[0030] 6) Add chloramphenicol monoclonal antibody and react at room temperature for 2-4 hours;

[0031] 7) Add an equal volume of 2% BSA and continue the reaction for 2-4h;

[0032] 8) Centrifuge at 10000rpm for 10min, discard the supernatant;

[0033] 9) Add 20 mL of particle resuspension solution (10 mM Tris-HCl, 1% skimmed milk powder, 5% trehalose), resuspend the particles, and set aside.

Embodiment 2

[0034] The preparation of embodiment 2 reaction cell

[0035] 1) According to the amount of 100 μL / well, add the fluorescent nanoparticle-chloramphenicol antibody conjugate to the 96-well microplate;

[0036] 2) Freeze at -80°C for 24 hours;

[0037] 3) Fluorescent nanoparticles-chloramphenicol antibody conjugates in freeze-dried wells;

[0038] 4) Store the lyophilized fluorescent nanoparticle-chloramphenicol antibody conjugate in a dry environment.

Embodiment 3

[0039] The preparation of embodiment 3 immunochromatography test strips

[0040] Preparation of the sample pad: cut the cellulose membrane into strips with a size of 1.5×30cm, and set aside;

[0041] Preparation of sample pad: Cut nitrocellulose membrane into 2.5×30cm strips, spray chloramphenicol-BSA conjugate (detection line) and goat anti-mouse secondary antibody (quality control line) on different positions of NC membrane , Fluorescent nanoparticles (reference line), dry at room temperature;

[0042] Preparation of absorbent pad: Cut the absorbent paper into 3.5×30cm strips for later use;

[0043] Assembly of test strips: Paste absorbent pads, nitrocellulose membranes, and sample pads on the bottom plate in sequence, cut into 5mm-wide test strips, and store in a dry place at 2-8°C.

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PUM

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Abstract

The invention belongs to the technical field of biomedicine and discloses an immunochromatographic device and a detection method using the same. The immunochromatographic device is composed of an immunochromatographic test strip and a reaction tank. In the immunochromatographic device, quick, ultrasensitive and quantitative detection of a target substance can be achieved by virtue of time distinguishing detection equipment by taking a rare-earth fluorescence nanoparticle as a marker. Compared with the traditional immunochromatographic test strip, the immunochromatographic device has high sensitivity. The device can be widely applied to quick, qualitative and quantitative detection items in the fields of biology, medicine, food safety and the like.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an immunochromatography detection device and a detection method for qualitatively and quantitatively detecting the content of a target substance in a sample. Background technique [0002] Immunochromatography technology is a simple and rapid detection technology in the early 1980s. Immunochromatography test strips made by this technology usually consist of the following parts: backing and sample pads pasted on the backing, binding pads, cellulose films, absorbent pads, etc. During detection, the sample is dropped onto the sample pad, and the sample will migrate on the chromatographic strip through capillary action. During the migration process, it will specifically react with the marker on the binding pad to generate an immune complex, which continues to Migrate and specifically combine with the corresponding antigen / antibody in the detection area on the cellulose membrane...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/533G01N33/531
Inventor 汤凌霄李金峰付辉王西丽
Owner 汤凌霄
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