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Fluorescent immunochromatography test strip for detecting furazolidone metabolites as well as preparation and application

A technique of fluorescence immunochromatography and furazolidone, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of cumbersome detection process, complicated operation, and expensive instruments, and achieves improved sensitivity, high quantum yield, and uniform particle size. Effect

Inactive Publication Date: 2016-10-26
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

HPLC has the characteristics of high detection accuracy and low false positive rate, but the instrument is expensive, the operation is cumbersome, time-consuming, expensive detection costs, high professional requirements for testing personnel, and complicated sample pretreatment; HPLC-MS and HPLC-MS / MS detection is accurate and repeatable, but the detection equipment is expensive and the operation is complicated, which is not conducive to wide-scale promotion and use; ELISA began to be used in the field of food safety detection in my country in the 1990s. This method is based on the specific reaction of antigen and antibody to detect The sample to be tested has high detection sensitivity and good specificity, which promotes the development of food safety detection technology
However, there are many experimental steps, cumbersome detection process, and poor reagent stability caused by enzyme catalysis; GICA is a relatively commonly used method in immunoassay methods, which has the advantages of time saving, high efficiency, and low cost, and the test results have good reproducibility and coefficient of variation. Small, very suitable for on-site detection and mass screening, but still has the disadvantages of insufficient indicator sensitivity and difficult preservation of test results

Method used

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  • Fluorescent immunochromatography test strip for detecting furazolidone metabolites as well as preparation and application

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Preparation of Aminated Water-Soluble Fluorescent Quantum Dots

[0041] 31.9mg Te powder, 28.5mg NaBH 4Add it into a small glass bottle with a self-made white rubber cover, blow it with nitrogen to deoxygenate, inject 5mL of anaerobic water with a syringe after half an hour, stir vigorously at room temperature for 1 hour, let it stand in an ice bath at 4°C for 10 minutes, and blow it with nitrogen during the whole process Carry out anaerobic protection, finally obtain the lavender NaHTe aqueous solution; Reaction simultaneously, 142.75mgCdCl 2 2.5H 2 O and 245ml of anaerobic water were added to a 250ml three-neck bottle, stirred magnetically, and then added to 0.1g polyethyleneimine (PEI) / 3mL ethylene glycol solution dissolved by ultrasound after the white solid was completely dissolved, and stirred for about 10 minutes , use 1mol / L NaOH solution to adjust the pH value to keep it alkaline, pass nitrogen gas, heat up to 105°C, after the pressure and temperature are sta...

Embodiment 2

[0043] Purification of fluorescent quantum dots

[0044] Quantum dots are purified by acetone precipitation and centrifugation. The specific operation is as follows: Take 20mL of CdTe QDs aqueous solution in a 50mL centrifuge tube, add an equal amount of acetone solution, let stand for 10min, and then centrifuge at 14000r / min at 4°C for 30min to remove colorless Supernatant, then add the same amount of acetone solution as the remaining solution, centrifuge, remove the colorless supernatant, repeat the above steps 1 to 2 times until a red precipitate, that is, red solid quantum dots is obtained, and the red solid quantum dots should be protected from light After drying, it was dissolved in 2mL PBS (0.01mol / L pH=7.4) and stored in the dark at room temperature to obtain the purified CdTe QDs PBS solution.

Embodiment 3

[0046] Preparation of Fluorescent Probes for AOZ Monoclonal Antibody Labeled with Fluorescent Nanoparticles

[0047] Take 2 mL of the purified CdTe QDs PBS solution in Example 2 above, add 120 ul MES buffer solution and 280 ul ultrapure aqueous solution, mix well, add 0.04 mg EDC for cross-linking, stir magnetically at room temperature for 15 min; add 20 ul aminocaproic acid, stir magnetically at room temperature for 30 min Add 1mL MES buffer, wash by centrifugation at 25000rpm for 30min, discard the supernatant, add 200ul MES buffer to dissolve the precipitate; add 0.04mg NHS and 0.02mg EDC, stir and mix at room temperature for 15min; add 1mL MES buffer, wash by centrifugation for 30min, discard Supernatant; add 400ul cross-linking buffer (0.05mol / L, boric acid buffer solution with pH=8.0), dissolve the precipitate, add AOZ monoclonal antibody (AOZ-mAb), stir magnetically at room temperature for 50min; add 80ul Tris-HCl (0.5 mol / L, pH=8.0) to terminate the reaction, add 50ul ...

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Abstract

The invention relates to a fluorescent immunochromatography test strip for detecting furazolidone metabolites as well as preparation and an application of the test strip. The fluorescent immunochromatography test strip comprises a bottom plate, a sample pad, a bonding pad, a chromatography membrane and a water absorption pad, wherein a detection line and a quality control line are arranged on the chromatography membrane; a fluorescent probe of a fluorescent nanoparticle marked AOZ (3-amina-2-oxazolldinone) monoclonal antibody is fixed on the bonding pad; the detecting line is coated with an antigen AOZ-BSA, and the quality control line is coated with a sheep anti-mouse IgG antibody. Compared with the prior art, according to the characteristic of high sensitivity of an amino-modified fluorescent CdTe quantum dot, the fluorescent probe of the marked AOZ monoclonal antibody is prepared, the AOZ fluorescent immunochromatography test strip is prepared and can be used for performing qualitative detection, semi-quantitative detection and auxiliary quantitative detection for a fluorescent reading instrument, so that the detection sensitivity is improved greatly, and the test strip is suitable for AOZ on-site detection and provides higher-value reference for detection of other pesticide residues and antibiotics.

Description

technical field [0001] The invention relates to a method for rapidly detecting furazolidone metabolites, in particular to a fluorescent immunochromatographic test strip for detecting furazolidone metabolites and its preparation and application, belonging to the technical field of fluorescent immunodetection. Background technique [0002] Furazolidone (Furazolidone), also known as furazolidone, chemically named 3-(5-nitrofurfural amino)-2-oxazolidinone, is a broad-spectrum nitrofuran antibiotic for common Gram-negative Bacteria and positive bacteria have inhibitory effect, and the other three (nitrofuran, furaltadone, nitrofurantoin) nitrofuran drugs are used for the prevention and treatment of dysentery, enteritis, gastric ulcer and other gastrointestinal diseases caused by bacteria and protozoa. Widely used in livestock and poultry and aquaculture. [0003] In the 1990s, it was discovered that furazolidone and its metabolite 3-amino-2-oxazolidinone (AOZ) can be absorbed by...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/543G01N33/558G01N33/577
CPCG01N33/533G01N33/54346G01N33/558G01N33/577
Inventor 魏新林王元凤余超徐乃丰汪艳姣徐凤
Owner SHANGHAI NORMAL UNIVERSITY
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