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A fluorescent immunochromatographic test strip prepared with aggregation-induced luminescent fluorescent microspheres as a beacon carrier

A technology of aggregation-induced luminescence and fluorescence immunochromatography, which is applied in the fields of medical inspection and food safety detection, can solve the problems of preparing fluorescent microspheres from materials such as tetramethyl styrene-acid, and achieve good fluorescence characteristics and fluorescence signals. Strong, high detection sensitivity effect

Active Publication Date: 2020-07-14
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on the above situation, the inventor has researched and developed a kind of fluorescent microspheres prepared by using the aggregation-induced luminescent material tetramethyl phenylacrylate (TCBPE). At present, there is no relevant report on the preparation and application of tetramethyl styrene acid tetramethyl styrene in fluorescent microspheres.

Method used

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  • A fluorescent immunochromatographic test strip prepared with aggregation-induced luminescent fluorescent microspheres as a beacon carrier
  • A fluorescent immunochromatographic test strip prepared with aggregation-induced luminescent fluorescent microspheres as a beacon carrier
  • A fluorescent immunochromatographic test strip prepared with aggregation-induced luminescent fluorescent microspheres as a beacon carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of aggregation-induced luminescent fluorescent microspheres (preparation of TCBPE dyes)

[0037] 1. Configure the dye prefabricated solution: weigh TCBPE (TCBPE synthesis method is from the literature RigidifyingFluorescent Linkers by Metal-Organic Framework Formation for FluorescenceBlue Shift and Quantum YieldEnhancement Journal of the American Chemical Society 2014 136(23), reported in 8269-8276), PMMA and PMAO, dissolved in chloroform, so that the final concentrations of TCBPE, PMMA, and PMAO dissolved in chloroform were 30, 50, and 25 mg / mL, respectively;

[0038] 2. Preparation of microemulsion: suck 2400 μL of the above pre-prepared solution, disperse it in 12 mL of 0.25% SDS solution, under ice bath conditions, sonicate for 3 min with a ultrasonic crusher, and the emulsification power is 600 W;

[0039] 3. Solidify into spheres: put the above emulsion in a rotary evaporation container, rotate until there is no chloroform, centrifuge the co...

Embodiment 2

[0040] Example 2: Preparation of a competitive immunochromatographic test strip for the detection of 25-hydroxyvitamin D (25-OH-VD) prepared by using aggregation-induced luminescent fluorescent microspheres as beacon carriers

[0041] 1. Preparation process of immunochromatographic test strips

[0042] 1. Preparation of nitrocellulose membrane;

[0043] (1) Preparation of 25-OH-VD artificial antigen (25-OH-VD-OVA):

[0044] The coupling method is mixed acid anhydride method, the coupling protein is ovalbumin (OVA), and the coupling ratio is 1:80. After coupling, dialysis and purification are performed to obtain 25-OH-VD-OVA.

[0045] (2) Preparation of test line and quality control line:

[0046] Coating of 25-OH-VD-OVA conjugate and anti-mouse antibody onto nitrocellulose membrane: Dilute the concentration of 25-OH-VD-OVA conjugate with 0.01 M PBS (phosphate buffered saline) pH 7.5 The concentration of the diluted anti-mouse antibody was 1 mg / mL, the obtained solution was ...

Embodiment 3

[0061] Example 3: Preparation of a competitive immunochromatographic test strip for the detection of zearalenone using aggregation-induced luminescent fluorescent microspheres as beacon carriers

[0062] 1. Preparation process of immunochromatographic test strips

[0063] 1. Preparation of nitrocellulose membrane;

[0064] (1) Preparation of ZEN artificial antigen (ZEN-BSA):

[0065] The coupling method is mixed acid anhydride method, the coupling protein is bovine serum albumin (BSA), and the coupling ratio is 1:100. After coupling, dialysis and purification are performed to obtain ZEN-BSA.

[0066] (2) Preparation of test line and quality control line:

[0067] Coating of ZEN-BSA conjugate and anti-mouse antibody onto nitrocellulose membrane: The ZEN-BSA conjugate was diluted to a concentration of 6 mg / mL with 0.01 M PBS (phosphate buffered saline) pH 7.0, and the resulting solution was Spray on the membrane as the detection line; the concentration of the diluted anti-mou...

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Abstract

The invention provides a fluorescence immunochromatography test strip prepared by taking an aggregation-induced emission fluorescent microsphere as a beacon carrier. Filter paper, a sample cushion, anAIEFM (Aggregation-Induced Emission Fluorescent Microsphere) compound glass fiber cushion, a nitrocellulose membrane sprayed with a detection line and a quality control line, and water adsorption paper are overlapped and pasted to a bottom plate in sequence to prepare the fluorescence immunochromatography test strip, the fluorescence immunochromatography test strip is used for quantitatively detecting the concentration of an object to be tested in a sample, and the prepared test strip has the characteristics of being high in detection sensitivity.

Description

technical field [0001] The invention belongs to the fields of medical inspection and food safety inspection, and in particular relates to a fluorescent immunochromatographic test strip prepared by using aggregation-induced luminescent fluorescent microspheres as beacon carriers to quantitatively detect the concentration of an analyte in a sample. Background technique [0002] Immunochromatography is a detection method based on the specific reaction of antigen and antibody. It has the advantages of fast detection speed, good specificity, simple operation and low cost. Compared with other methods, it can better meet the needs of on-site large-scale detection. Therefore, In recent years, it has developed rapidly and is widely used in the fields of food safety, medical inspection and environmental pollutant monitoring. Among them, colloidal gold immunochromatography test strip is the most widely used immunochromatography product, but it has poor sensitivity and is susceptible to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/553
CPCG01N33/533G01N33/553
Inventor 赖卫华熊勇华魏华陈媛章钢刚许恒毅喻志标彭娟
Owner NANCHANG UNIV