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Immunochromatographic test strip for detecting malachite green (MG) and preparation process thereof

An immunochromatographic test paper and malachite green technology, which can be used in measuring devices, analytical materials, instruments, etc., can solve the problems of limiting superparamagnetic composite particles, long color development time, cumbersome and complicated problems, etc.

Inactive Publication Date: 2014-04-02
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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  • Immunochromatographic test strip for detecting malachite green (MG) and preparation process thereof
  • Immunochromatographic test strip for detecting malachite green (MG) and preparation process thereof
  • Immunochromatographic test strip for detecting malachite green (MG) and preparation process thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1, Preparation of Malachite Green Residue Magnetic Label Rapid Detection Test Paper

[0089] (1) Preparation of malachite green antibody labeled with superparamagnetic composite particles

[0090] 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 malachite green antibody.

[0091] The specific method is: 1) Take 2.5mg of the above-mentioned magnetic particles and wash them with 0.1M MES buffer (weigh 1.066gMES, 0.45g NaCl and dissolve them in 50ml pure water, adjust the pH to 4.7) and separate and enrich them with a 0.4T magnetic rack Afterwards, resuspend with 1ml of MES buffer solution with a concentration of 0.1M and a pH value of 4.7, and add 0.96mg (final concentration of 5mM) of 1-ethyl-(3-di...

Embodiment 2

[0107] Embodiment 2, the detection of the sensitivity, cross-reactivity and accuracy of the immunochromatographic test paper of malachite green residue.

[0108] 1. Determination of sensitivity

[0109] Weigh an appropriate amount of malachite green (Beijing Hengyuan Qitian Chemical Technology Research Institute, catalog number: 1245BW3811), and dissolve it in deionized water to prepare a standard stock solution of 100ug / ml. Dilute the standard solution prepared as 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10ug / L with 0.02M PBS (pH=7.4), add the immunoassay for detecting malachite green obtained by Example 1 respectively Chromatographic test paper, and read with a superparamagnetic resonance detector MAR (Magna BioSciences, 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 magnet...

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Abstract

The invention discloses an immunochromatographic test strip for detecting MG and a preparation process thereof. The immunochromatographic test strip for detecting MG provided in the invention comprises a sample pad containing MG antibodies marked by superparamagnetic composite particles, a cellulose nitrate membrane connected with one end of the sample pad and an absorbent connected with another end of the cellulose nitrate membrane; the cellulose nitrate membrane is coated by mutually separated detection lines and quality control lines, wherein, the detection lines contain MG antigens, and the quality control lines contain antiantibodies which can specifically bind to the MG antibodies. Experiments in the invention prove that the test strip provided in the invention has the advantages of high sensitivity, strong specificity, rapidness, simpleness and capability of realizing objective detection when is used for detecting MG.

Description

technical field [0001] The invention relates to a detection reagent for malachite green residues, in particular to an immunochromatographic test paper for detecting malachite green and a preparation method thereof. Background technique [0002] Malachite green (malachite green, MG), also known as basic green, base block green, malachite green, aniline green, Victoria green or Chinese green, belongs to triphenylmethane dyes and has been widely used in the prevention and treatment of various diseases. Saprolegniasis, gill mold and melon worm disease of aquatic animals. However, malachite green and its metabolite colorless malachite green will produce high residues in aquatic products. After human consumption, it is easy to cause side effects such as carcinogenicity, teratogenicity and mutagenicity. The European Union, the United States and some East Asian countries have banned the use of malachite green as an antifungal drug for fish. In May 2002, my country also included ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/577
Inventor 马岚袁航吴峰
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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