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Immunochromatographic quantitative test reagent based on near-infrared fluorescent marker

An immunochromatographic test strip and fluorescent labeling technology, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of inaccurate quantification and low sensitivity of immunochromatographic reagents.

Active Publication Date: 2012-09-19
BEIJING RUNBO FUDE BIOLOGICAL TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention overcomes the problems of low sensitivity and inaccurate quantification of immunochromatography reagents using colloidal gold particles as markers, and establishes a quantitative detection reagent for immunochromatography based on near-infrared fluorescent markers and a new detection method. Quantitative detection reagent package detection test strip and preparation method of test strip

Method used

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  • Immunochromatographic quantitative test reagent based on near-infrared fluorescent marker
  • Immunochromatographic quantitative test reagent based on near-infrared fluorescent marker
  • Immunochromatographic quantitative test reagent based on near-infrared fluorescent marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Sandwich method detects human hemoglobin

[0030] 1) Infrared fluorescence labeled hemoglobin monoclonal antibody and chicken IgY.

[0031] Hemoglobin monoclonal antibody (British Abcam Company) and chicken IgY antibody (UK Abcam Company) were dialyzed with PBS, and the antibody was mixed with the fluorescent dye according to the ratio of 7 μl NHS-activated Dylight800 near-infrared fluorescent dye (American Thermo Company) to label 1 mg hemoglobin monoclonal antibody, After mixing evenly, react at room temperature in the dark for 1 hour. After the reaction, put the labeled product into a dialysis bag with a pore size of 10K, and dialyze with PBS at 4°C overnight to remove free fluorescent dye. The solution was added with a final concentration of 1% BSA and 0.1% Tween20, sodium azide 0.1‰, and stored at 4°C.

[0032] 2) Make bonding pads

[0033] Select the glass cellulose strip as the solid phase material of the binding pad, spray two kinds of near-infr...

Embodiment 2

[0049] Embodiment 2: Sandwich method detects HIV antibody

[0050] 1) Near-infrared fluorescent labeling of biological macromolecules

[0051] The genetically engineered recombinant HIV-1gp41 (Immune Technology Company) and HIV-2gp36 (Immune Technology Company) antigens were dialyzed with PBS, and the ratio of 1 mg of recombinant antigens labeled with 7 μl of NHS-activated near-infrared fluorescent dye Dylight 800 (Thermo Electric Company of the United States) was adjusted to two After mixing evenly, react at room temperature in the dark for 1 hour. After the reaction, put the labeled product into a dialysis bag with a pore size of 10K, and dialyze with PBS at 4°C overnight to remove free fluorescent dye. The solution was added with a final concentration of 1% BSA and 0.1% Tween20, sodium azide 0.1‰, and stored at 4°C. Select glass cellulose tape as the solid-phase material of the binding pad, spray the near-infrared fluorescent dye-biomolecule solution on the strip, and air...

Embodiment 3

[0067] Embodiment 3: Indirect method detects HIV antibody

[0068] 1) Dylight800 labeled detection antigen and quality control antibody

[0069] The goat anti-human IgG antibody (purchased from Abcam) was dialyzed with PBS, and the two were mixed according to the ratio of 7 μl of NHS-activated Dylight800-labeled 1 mg antibody. After mixing evenly, react in the dark at room temperature for 1 hour. After the reaction, put the labeled product into a dialysis bag with a pore size of 10K, and dialyze with PBS at 4°C overnight to remove free fluorescent dye. The solution was added with a final concentration of 1% BSA and 0.1% Tween20, sodium azide 0.1‰, and stored at 4°C.

[0070] 2) Make bonding pads

[0071] Select glass cellulose tape as the solid-phase material of the binding pad, spray the near-infrared fluorescent dye-biomolecule solution on the strip, and air-dry the strip at room temperature.

[0072] 3) Preparation of sample pad

[0073] Select the cellulose membrane st...

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Abstract

The invention relates to an immunochromatographic quantitative test reagent based on a near-infrared fluorescent marker. A near-infrared fluorescent molecule is adopted as a marker, and the immunochromatographic technology is adopted to prepare a near-infrared fluorescent immunochromatographic test strip. In the test process, a conventional near-infrared test instrument is used for respectively scanning a quality control line and a sample line through near-infrared rays, and after being corrected by utilizing the fluorescent strength of the quality control line, the fluorescent strength of a detection line is substituted in a standard curved line in a fluorescent analysis instrument, so that the concentration of an object to be tested in a test specimen can be analyzed. The reagent can be applied to the detection of microorganism, the food safety detection, the drug detection and the quick detection of dangerous chemicals. The immunochromatographic quantitative test reagent has characteristics of high sensitivity, accuracy in quantization and convenience in operation.

Description

technical field [0001] The invention establishes an immunochromatographic quantitative detection reagent based on a near-infrared fluorescent marker. The preparation method includes near-infrared fluorescence detection test strips and corresponding reagents. The present invention uses near-infrared fluorescent molecules as markers, adopts immunochromatographic technology to prepare near-infrared fluorescent immunochromatographic test strips, and then forms a detection card comprising sample pad / glass fiber membrane / nitrocellulose membrane and absorbent paper, wherein nitric acid A test line and a quality control line are fixed on the cellulose membrane. In the detection process, the near-infrared light is used to scan the quality control line and the sample line respectively, and the fluorescent molecules are excited to emit light. The emitted fluorescence is converted into a digital signal by a filter and a photomultiplier tube, and the detection line is corrected by the flu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558
Inventor 李志刚陈唯军
Owner BEIJING RUNBO FUDE BIOLOGICAL TECH DEV
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