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Immunofluorescence test strip based on phosphorescent technology, and preparation method and application thereof

A technology of immunofluorescence and test strips, applied in biological testing, material inspection products, measuring devices, etc., can solve problems such as cytotoxicity, flickering fluorescence, interference of cell physiological processes, etc., and achieve sensitive qualitative and quantitative detection and analysis Effect

Inactive Publication Date: 2013-01-09
SHENZHEN AIRUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when quantum dots are used as fluorescent markers, there are still some problems, such as aggregation in solution, stability after biomarking, scintillation fluorescence, background fluorescence interference of complex biological samples, and potential cytotoxicity. and interference with cellular physiological processes, etc.

Method used

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  • Immunofluorescence test strip based on phosphorescent technology, and preparation method and application thereof
  • Immunofluorescence test strip based on phosphorescent technology, and preparation method and application thereof
  • Immunofluorescence test strip based on phosphorescent technology, and preparation method and application thereof

Examples

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Effect test

preparation example Construction

[0051] ① Preparation of phosphorescent material markers:

[0052] Dilute biologically active molecules with buffer solution, add phosphorescent material solution, stir well, react at room temperature for at least 1 hour, separate and purify with G25 gel column, collect markers, and dilute with phosphate buffer Store after mixing;

[0053] ② Preparation of sample pad:

[0054] Use cellulose membrane as the solid phase material of the sample pad, cut into strips, and use 0.01% to 0.5% polyethylene glycol, 1% to 5% bovine serum albumin, and 0.01% to 0.05% surfactant in 0.01 to 0.3 Soak in phosphate buffer solution, the pH value of the buffer solution is 7.2-7.6. After soaking, put the sample pad in the drying oven to dry, take it out, and vacuum seal it for later use;

[0055] ③Preparation of the conjugate pad:

[0056] Use glass cellulose membrane as the solid phase material of the conjugate pad, cut into strips, and use 1% to 5% bovine serum albumin, 0.1 to 2% polyethylene g...

Embodiment 1

[0077] Example 1: Detection of hepatitis B virus surface antigen HBsAg by double-antibody sandwich method

[0078] The method for preparing quantitatively detecting hepatitis B virus surface antigen immunochromatographic test strips comprises the following steps:

[0079] 1. Preparation of anti-HBs labeled with phosphorescent luminescent material:

[0080] Dilute anti-HBs monoclonal antibody B and goat anti-rabbit IgG antibody with 0.1M pH9.6 sodium bicarbonate-sodium carbonate solution to 1mg / ml respectively, take 5ml of antibody solution, add 30mg of phosphorescent material metalloporphyrin to dissolve solution, stir well, incubate at room temperature for 1 hour, and mix every 15 minutes. Finally, the G25 gel column was used for separation and purification, and the labeled metalloporphyrin-labeled anti-HBs antibody B and goat anti-rabbit IgG antibody were collected and used to contain 0.1% polyethylene glycol, 2% bovine serum albumin, Dilute with 0.01M pH7.2 phosphate buff...

Embodiment 2

[0108] Example 2: Detection of HIV Antibody by Double Antigen Sandwich Method

[0109] The preparation method of qualitative detection HIV antibody immunochromatographic test strip comprises the following steps:

[0110] 1. Preparation of HIV antigen labeled with phosphorescent material:

[0111] Dilute HIV antigen B and goat anti-rabbit IgG antibody to 1mg / ml with 0.05M pH9.6 sodium bicarbonate-sodium carbonate solution respectively, take 5ml antibody solution respectively, add 30mg phosphorescent material metalloporphyrin solution respectively, stir Mix well and incubate at room temperature for 1 hour, mixing every 15 minutes. Finally, use a G25 gel column for separation and purification, collect the marked metalloporphyrin-labeled HIV antigen B and goat anti-rabbit IgG antibody, and use 0.1% polyethylene glycol, 2% bovine serum albumin, 2 % sucrose, 0.05% surfactant, diluted with 0.01M pH7.2 phosphate buffer solution, packed in sealed reagent bottles, and stored at 2-8°C....

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Abstract

The invention provides an immunofluorescence test strip based on a phosphorescent technology, and a preparation method and application thereof. A phosphorescent material, namely pt-porphyrin / pd-porphyrin, is used as a biomarker of the test strip, a result is shown in a mode of infrared light signals under illumination of green light, and can be identified by an instrument, so that quantitative detection for a detected object is realized. The test strip comprises a sample pad, a conjugate pad, an analyzing membrane, a water-sucking pad and a liner. A phosphorescent material marker is fixed on the conjugate pad, and a detecting line and a quality control line are fixed on the analyzing membrane. The invention also discloses a preparation method for the test strip and application of the test strip in quantitative detection of biological samples. According to difference of immunoreaction modes of an object to be detected, the test strip includes a sandwich method mode, a competition method mode, an indirect method mode and a capture method mode; and according to difference of properties of the detected object, different objects to be detected in the sample can be quickly and sensitively detected and analyzed qualitatively and quantitatively by using different detection modes.

Description

technical field [0001] The invention relates to an immunofluorescence test strip based on phosphorescence technology, a preparation method and application thereof, and belongs to the technical field of immunological test strips. Background technique [0002] Immunoassay is based on the immune binding reaction between antigen and corresponding antibody, that is, using antibody (or antigen) as a selective reagent to determine antigen, hapten (or antibody). The reaction between antibody and antigen has extremely high specificity, so no or only simple pretreatment is required for immunoassay samples. Antibody-antigen complexes have high stability, and their stability constants are generally 10 9 , even up to 10 15 . Unfortunately, although antibody-antigen binding can display certain catalytic properties, from the perspective of analytical testing, the immune response lacks the analytical signal contrast of using highly sensitive reagents, making it difficult to detect direct...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577G01N33/531G01N33/571G01N33/576G01N33/74G01N33/569
Inventor 谢爱武
Owner SHENZHEN AIRUI BIO TECH
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