Fluorescent probe, method for detecting tetracycline and application

A fluorescent probe, tetracycline technology, used in measurement devices, biological tests, material inspection products, etc., can solve the complex cross-linking process of antibodies and signal carriers, etc., to maintain fluorescence signal intensity, high fluorescence signal intensity, high sensitivity Effect

Active Publication Date: 2020-08-18
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the defects and deficiencies in the prior art, in order to solve the problem of complex cross-linking process between antibodies and signal carriers, the present invention constructs a fluorescent carrier to enrich signal substances. The fluorescent probe has high fluorescence intensity, stable chemical / physical properties and Combining antibodies does not require complex cross-linking agents, which is of great significance and application value for monitoring the residual tetracycline in food, including a fluorescent probe, a method for detecting tetracycline and its application

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  • Fluorescent probe, method for detecting tetracycline and application
  • Fluorescent probe, method for detecting tetracycline and application
  • Fluorescent probe, method for detecting tetracycline and application

Examples

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Embodiment 1

[0045] Following the above technical scheme, this example presents the fluorescent probe AlNPs-Ab and its preparation method, including preparing amino-functionalized aluminum nanosheets as signal carriers by hydrothermal method, and then adding tetracycline monoclonal antibody for adsorption. Include the following steps:

[0046] (1) Preparation of amino-functionalized aluminum nanosheets (AlNPs): Mix aluminum chloride and diaminoterephthalic acid for reaction, then add urea solution, stir and transfer the mixture to a high-pressure reactor, and then wash and centrifuge. have to;

[0047] Specifically: the AlCl 3 and diaminoterephthalic acid (NH 2 -BDC) was added into 15mL deionized water at a concentration ratio of 1:1 and mixed completely, and the reaction was continued for 30min. Subsequently, urea solution (0.96 mol / L, 5 mL) was added dropwise under vigorous stirring, and stirring was continued for 30 min. Afterwards, the mixture was transferred to an autoclave and re...

Embodiment 2

[0052] This embodiment provides a high-sensitivity immunochromatographic test strip for rapid detection of tetracycline, including a liner, a nitrocellulose membrane is pasted on the liner, one end of the nitrocellulose membrane is covered with an absorbent pad, and the other end of the nitrocellulose membrane is The sample pad and the binding pad are covered in sequence, and the detection line and the control line are arranged laterally on the non-covered surface of the nitrocellulose membrane, and the binding pad and the sample pad are respectively sealed with a blocking solution.

[0053] The preparation method of the nitrocellulose membrane includes: 1 mg / mL tetracycline-bovine serum albumin conjugate is coated on the detection line at a streaking rate of 1 μL / cm to obtain a detection line, and 1 mg / mL goat anti-mouse immunoglobulin is coated with 1 μL The scribe rate of / cm was coated on the control line as the control line; then dried at 37°C for use.

[0054] Preparatio...

Embodiment 3

[0058] Embodiment 3: the sensitivity determination of the test strip of rapid detection tetracycline

[0059] Performance evaluation of immunochromatographic test strips for the rapid detection of tetracycline as described above.

[0060] Detection process: Dissolve tetracycline standard substance in ultrapure water, serially dilute to make the test solutions with different concentrations ranging from 0 to 12.0ng / mL, and ultrapure water is used as blank control. Mix and incubate 0.5 μL of AlNPs-Ab probe with 100 μL tetracycline standard solution, then immerse the sample pad of the test strip into 100 μL test solution, and the mixture migrates to the absorbent pad through capillary action. After 15 min of reaction, the strips were scanned using a fluorescent strip reader to obtain fluorescence intensity (FI). Use the FI ratio (FI T / FI C ) for quantitative analysis, which can not only offset the variability between test strips, but also minimize the environmental factors aff...

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Abstract

The invention discloses a fluorescent probe, a method for detecting tetracycline and an application of the fluorescent probe. The probe comprises a tetracycline monoclonal antibody and a signal carrier, wherein the signal carrier is an amino-functionalized aluminum nanosheet, and the particle size of the amino-functionalized aluminum nanosheet is 520-570nm. According to the invention, the probe isconstructed by a fluorescent metal organic framework material in immunochromatographic test strip detection for the first time, and a simple, novel, sensitive and rapid analysis system is developed for tetracycline detection of meat, milk, honey and other actual samples. The lowest detection limit of the provided test strip on tetracycline is 0.0516 ng / mL, and the value of the lowest detection limit is less than that reported by many other literatures.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a fluorescent probe, a method for detecting tetracycline and its application, in particular to a probe which is marked with a fluorescent metal organic framework material and enriched with a monoclonal antibody, and which is detected by a test strip using the probe A method for tetracycline, and its use for rapid and sensitive detection of tetracycline. Background technique [0002] Tetracycline (TET), as a broad-spectrum antibiotic, has been widely used as a feed additive to enter the food chain. Long-term intake of food containing tetracycline residues can cause allergic reactions in humans, gastrointestinal discomfort, and even liver toxicity. Immunochromatographic test strips (LFIA) are the most common point-of-care diagnostic tools for the detection of various mycotoxins, proteins, antibiotics, heavy metals, etc. [0003] The current methods for detecting tetracyclines mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/577G01N33/558G01N33/53G01N33/52
CPCG01N33/582G01N33/577G01N33/558G01N33/5308G01N33/52Y02A50/30
Inventor 王丽白菲儿补彤张萌田永明孙新玉贾珮李睿赵爽何坤益
Owner NORTHWEST A & F UNIV
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