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Biological probe, test strip for detecting furazolidone and applications thereof

A furazolidone and biological probe technology, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of not being able to meet the detection requirements well, the antibody coupling conditions are not well controlled, and the preparation process of labeling materials is complicated. , to achieve the effect of simple operation, less antibody dosage and high sensitivity

Inactive Publication Date: 2017-12-26
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In recent years, research has been devoted to replacing markers to improve detection sensitivity, but new marker materials (fluorescent microspheres, quantum dots, magnetic beads, graphene, etc.) are usually complicated to prepare, and the conditions for coupling with antibodies are not easy to control, and Some materials require signal readout equipment, which are not suitable for on-site detection
On the other hand, traditional colloidal gold test strips cannot achieve high sensitivity, and cannot well meet the detection requirements of the current stricter furazolidone limit standard

Method used

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  • Biological probe, test strip for detecting furazolidone and applications thereof
  • Biological probe, test strip for detecting furazolidone and applications thereof
  • Biological probe, test strip for detecting furazolidone and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Preparation method of furazolidone monoclonal antibody

[0040] The preparation method of furazolidone monoclonal antibody comprises the following steps:

[0041] 1. Ascites collection:

[0042] Furazolidone cells were cultured in advance, and when the cells grew to a good state, the cell line was injected into BALB / c mice that had been treated with liquid paraffin in advance, and the ascites of the mice was collected when the abdomen of the mice was obviously enlarged.

[0043] 2. Antibody purification:

[0044] The antibody was purified by caprylic acid-ammonium sulfate method, and the specific operation was as follows: filter the ascites with double-layer filter paper, centrifuge at 12000 r / min for 15 min at 4°C, and absorb the supernatant. The ascites supernatant obtained was mixed with 4 times the volume of acetate buffer, and n-octanoic acid 33 μL / mL ascites was slowly added under stirring, mixed at room temperature for 30 minutes, and left standing at 4°C for 2...

Embodiment 2

[0051] Nano gold solution is prepared by sodium citrate reduction method. The specific method is: take 96mL of distilled water in the Erlenmeyer flask, after heating and boiling, add 1mL of HAuCl successively under the state of heating and magnetic stirring 4 and 4mL trisodium citrate solution, observe the color change, from blue to purple to red, after the solution turns red, continue to heat for 10min under stirring. After the solution was cooled at room temperature, the volume was adjusted to 100 mL with distilled water in a volumetric flask, and placed in a 4°C refrigerator for later use. The particle size of nano gold is 15-20nm.

[0052] The preparation method of detection probe comprises:

[0053] Measure 50.0mL of nano-gold solution, adjust the pH of the solution to 5.5 with 0.1mol / L potassium carbonate solution; slowly add 0.35mL of 1mg / mL furazolidone monoclonal antibody solution while stirring, and continue stirring for 30min; add 10% Bovine serum albumin solutio...

Embodiment 3

[0058] Preparation of immunochromatographic test strips for rapid detection of furazolidone metabolites

[0059] High-sensitivity immunochromatographic test strips for the rapid detection of furazolidone metabolites, see figure 1 , which is provided with a liner, and the liner is attached with a water-absorbing pad, a nitrocellulose membrane, a binding pad containing two gold-labeled antibodies (gold-labeled monoclonal antibody and gold-labeled secondary antibody), and a sample pad in sequence from top to bottom. Wherein, a detection line is arranged laterally on the nitrocellulose membrane, and the detection line is coated with furazolidone metabolite-bovine serum albumin conjugate (CPAOZ-BSA).

[0060] The preparation method of rapid detection of furazolidone metabolites high-sensitivity immunochromatographic test strips comprises the following steps:

[0061] 1) Preparation of absorbent pad

[0062] Cut the absorbent paper to a length of 18mm and a width of 3mm to obtain ...

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Abstract

The invention discloses a biological probe, a test strip for detecting furazolidone and applications thereof. A nano gold labeled anti-furazolidone antibody is taken as the detection probe. A nano gold labeled goat-anti-mouse immune globulin G is taken as the enhancing probe. Two probes are sprayed on a hydrophobic pad. When a sample solution flows, two probes are eluted from the hydrophobic pad; the gold of two probes is aggregated through the combination of antibody and second antibody, a complicated network structure is formed, the gold flows forward under the capillary action, a clear red strip is formed in the T line, a signal amplifying effect is achieved, at the same time, the using amount of antibody is obviously reduced, more fierce competing reactions are triggered, and the sensitivity is improved.

Description

technical field [0001] The invention belongs to the field of biological detection, and specifically relates to a biological probe, a test strip for detecting furazolidone and an application thereof. Background technique [0002] Furazolidone is one of the most commonly used nitrofuran antibiotics (others mainly include nitrofurazone, furaltadone and nitrofurantoin). It is rapidly metabolized in the body and has a short half-life, so the parent drug cannot exist stably. However, the metabolite 3-amino-2-oxazolone (AOZ) of furazolidone can be tightly combined with the protein in the tissue, and exists stably in the body in the form of a bound state, and the longer the residual time, the toxicity of this type of drug stronger. [0003] Because furazolidone antibiotics are cheap and have killing effects on bacteria, protozoa and other pathogens and fungi, they are widely used in livestock and aquatic products to treat and prevent various gastrointestinal infections caused by ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/532G01N33/15G01N33/64
CPCG01N33/558G01N33/532G01N33/577G01N33/64G01N33/94G01N33/9446
Inventor 张道宏窦磊娜王建龙补彤黄琼闫灵芝赵兵欣赵东阳
Owner NORTHWEST A & F UNIV
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