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Immunochromatography test paper for detecting ractopamine and preparation method thereof

An immunochromatographic test paper and ractopamine technology, which is applied in the field of immunochromatographic test paper for detecting ractopamine and its preparation, and can solve the problems of easy polymerization of magnetic beads, slow swimming time, long color development time, etc.

Inactive Publication Date: 2014-03-19
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these preparation and modification methods are often cumbersome and complicated, and the obtained superparamagnetic composite particles cannot meet the requirements of LFIAs in terms of size, biocompatibility, saturation magnetic strength, external magnetic field response speed, stability, and labeling efficiency. The size is mostly between 200 and 300nm. Due to the large size of the magnetic beads, the swimming time on the test paper is slow and the color development time is long; and the particles that are too small cannot provide sufficient magnetic resonance signals; in addition, there are biological Unstable compatibility and easy aggregation of magnetic beads; these shortcomings limit the application of superparamagnetic composite particles in LFIAs

Method used

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  • Immunochromatography test paper for detecting ractopamine and preparation method thereof
  • Immunochromatography test paper for detecting ractopamine and preparation method thereof
  • Immunochromatography test paper for detecting ractopamine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1, Preparation of magnetically labeled rapid detection test paper for ractopamine residues

[0090] (1) Preparation of ractopamine antibody labeled with superparamagnetic composite particles

[0091] The particle size is 100nm, the magnetic saturation intensity is 40emu / g, the corresponding external magnetic field response speed is 20 seconds, and the superparamagnetic composite particle (superparamagnetic Fe 3 o 4 Nanoparticles) (purchased from Shenzhen Thales Technology Co., Ltd., catalog number MP-2) labeled ractopamine antibody.

[0092] The specific method is:

[0093] 1) Take 2.5mg of the above-mentioned magnetic particles, wash them with 0.1M MES buffer (weigh 1.066g MES, 0.45g NaCl and dissolve them in 50ml pure water, adjust the pH to 4.7), separate and enrich them with a 0.4T magnetic rack, and use 1ml of MES buffer with a concentration of 0.1M and a pH value of 4.7 was resuspended, and 0.96mg (final concentration of 5mM) of 1-ethyl-(3-dimethylamino...

Embodiment 2

[0108] Embodiment 2, the detection of the immunochromatographic test paper sensitivity, cross-reactivity and accuracy of ractopamine residue

[0109] 1. Determination of sensitivity

[0110] Weigh an appropriate amount of ractopamine (Beijing Hengyuan Qitian Chemical Technology Research Institute, catalog number: 30230CDCT-C16805000), and prepare a 100ug / ml standard stock solution with methanol. Dilute the standard solution prepared as 0, 0.005, 0.01, 0.05, 0.1, 1, 5, 10ug / L with 0.02M PBS (pH=7.4), add the immunochromatographic test paper for detecting ractopamine obtained by Example 1 respectively , and read using superparamagnetic resonance detector MAR (MagnaBioSciences, 8094-101-01 & 8094-101-02). Detection steps: Return the sample to be tested to room temperature (25°C) before detection, use a precise pipette to take 100 μl of the sample to be tested and drop it vertically and slowly into the sample loading end of the magnetic test strip, and then drop into 50ul of wash...

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Abstract

The invention discloses immunochromatography test paper for detecting ractopamine and a preparation method thereof. The immunochromatography test paper for detecting ractopamine comprises a sample pad, a cellulose nitrate film and a water absorbing pad, wherein the sample pad comprises a superparamagnetic compound particle marking ractopamine antibody; the cellulose nitrate film is connected to one end of the sample pad; the water absorbing pad is connected to the other end of the cellulose nitrate film; the cellulose nitrate film is wrapped by a detection line and a quality control line which are separated from each other; the detection line comprises a ractopamine antigen; and the quality control line comprises an anti-antibody which can be specially combined with the ractopamine antibody. As proved by an experiment, the test paper for detecting ractopamine, provided by the invention, has high sensitivity and high specificity, is rapid and convenient, and can be used for realizing objective detection.

Description

technical field [0001] The invention relates to a detection reagent for ractopamine residues, in particular to an immunochromatographic test paper for detecting ractopamine and a preparation method thereof. Background technique [0002] Ractopamine (RAC) is a β-stimulant, which has been illegally added to feed in recent years to increase the lean meat rate of fat animals and accelerate animal growth, because its additive dose is 5-10 times the therapeutic dose, so that High residues in animals can cause harm to consumers. At present, ractopamine colloidal gold test paper is mostly used as detection reagent for on-site screening, but it cannot meet the requirements well due to its low sensitivity. Therefore, it is necessary to develop a more accurate and rapid detection method in the field. [0003] Lateral flow immunochromatographic detection technology (LFIAs) based on antigen-antibody immunological reaction is a new technology developed in the early 1990s. Because of its ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/532G01N33/544
Inventor 马岚卢体康秦智锋袁航吴峰
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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