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Fluorescent microsphere immunochromatographic test strip for detecting metalaxyl and its preparation method and application

A technology of immunochromatographic test paper and fluorescent microspheres, applied in the field of metalaxyl detection, can solve the problem that there is no metalaxyl fluorescent microsphere immunochromatographic test strip, etc., and achieves easy popularization, cost reduction, and increased response The effect of sensitivity

Active Publication Date: 2021-08-17
ZHENGZHOU TOBACCO RES INST OF CNTC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Up to now, there is no metalaxyl fluorescent microsphere immunochromatographic test strips on the market

Method used

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  • Fluorescent microsphere immunochromatographic test strip for detecting metalaxyl and its preparation method and application
  • Fluorescent microsphere immunochromatographic test strip for detecting metalaxyl and its preparation method and application
  • Fluorescent microsphere immunochromatographic test strip for detecting metalaxyl and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Composition of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Metalaxyl Residues

[0041] see figure 1 , 2: The test strip is composed of a bottom plate, a sample binding pad, a nitrocellulose membrane and an absorbent pad.

[0042] The sample binding pad 1, nitrocellulose membrane 2 and water-absorbing pad 3 are overlapped and pasted on the bottom plate 6 in sequence, the end of the sample binding pad is connected to the beginning of the nitrocellulose membrane, and the end of the nitrocellulose membrane is connected to the water-absorbing pad. The beginning of the sample binding pad is aligned with the beginning of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom plate.

[0043] The nitrocellulose membrane is fixed with a detection area 4 and a quality control area 5, the detection area is sprayed with a metalaxyl hapten-carrier protein conjugate, and the quality control area is sprayed with ...

Embodiment 2

[0045] Example 2 Preparation of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Metalaxyl Residues

[0046] The preparation method of this test strip mainly comprises the following steps:

[0047] 1) Preparation of the sample binding pad: label the metalaxyl monoclonal antibody with commercially available fluorescent microspheres, and dilute it with a specific buffer system, soak the sample binding pad in the dilution buffer, and prepare it after vacuum freeze-drying ;

[0048] 2) Preparation of nitrocellulose membrane: Spray the metalaxyl hapten-carrier protein conjugate on the detection area of ​​the nitrocellulose membrane to make the detection area; spray goat anti-mouse anti-antibody onto the nitrocellulose membrane The scope of the quality control area above is made into the quality control area;

[0049] 3) Assembly and cutting: The sample binding pad embedded with fluorescent microsphere-labeled metalaxyl monoclonal antibody, the nitrocellulos...

Embodiment 3

[0098] Example 3 Application of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Metalaxyl Residues

[0099] 1. Tobacco sample pretreatment

[0100] Weigh 1.0±0.05 g crushed tobacco leaf sample into a 50 mL centrifuge tube, add 10 mL 50% methanol aqueous solution, vortex for 1 min, centrifuge at 3000 rpm for 5 min, take 100 μL supernatant and add 400 μL deionized water to dilute to be tested.

[0101] 2. Test with test strips

[0102] Draw 100 μL of the sample solution to be tested and drop it vertically into the sample hole of the test paper card. When the liquid flows, start timing and react for 10 minutes; insert the test paper card into the carrier of the KFT-100A fluorescence detector, and select the sample to be Check the items, press the "Start Test" button, the fluorescence detector will automatically scan the test paper card, and read or print the test results on the display screen of the instrument.

[0103] 3. Analysis of test results

[0...

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Abstract

The invention discloses a fluorescent microsphere immunochromatographic test strip for detecting metalaxyl residues, a preparation method and application thereof. The test strip includes a bottom plate and a sample binding pad, a nitrocellulose membrane, and a water-absorbing pad that are sequentially lapped and pasted on the bottom plate. The metalaxyl monoclonal antibody labeled with fluorescent microspheres is embedded on the sample binding pad, and the metalaxyl monoclonal antibody labeled with fluorescent microspheres is embedded on the nitrocellulose membrane. The detection area and the quality control area are fixed. The detection area is sprayed with metalaxyl hapten-carrier protein conjugate, and the quality control area is sprayed with goat anti-mouse anti-antibody. Metalaxyl monoclonal antibody is based on metalaxyl hapten-carrier protein conjugate. The carrier protein conjugate is prepared as an immunogen, and the metalaxyl hapten is obtained by a substitution reaction between metalaxyl and 4-formyl butyryl chloride under the catalysis of aluminum chloride. The invention also provides a preparation method of the test strip and a method of using the test strip to detect metalaxyl residues in tobacco and tobacco products. The test strip of the invention has the advantages of high sensitivity, rapid detection, convenient operation, economical and practical, and the like.

Description

technical field [0001] The invention belongs to the field of metalaxyl detection, and in particular relates to a fluorescent microsphere immunochromatographic test strip for detecting metalaxyl residues and its preparation method and application, which is especially suitable for the detection of metalaxyl residues in tobacco and tobacco products . Background technique [0002] Metalaxyl (Metalaxyl) is an amide systemic fungicide with preventive and therapeutic effects. It is used to prevent and control downy mildew and blight of melons, fruits and vegetables, potato early and late blight, tobacco black shank, etc. Although metalaxyl is not highly toxic to humans and animals, it belongs to systemic sterilization, and the residues in agricultural products and vegetables often exceed the standard seasonally. Therefore, the residue of metalaxyl in crops has also received high attention. GB 2763-2016 "National Food Safety Standard Maximum Residue Limits of Pesticides in Food" st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/58G01N33/533
CPCG01N33/533G01N33/558G01N33/582G01N33/585
Inventor 陈黎范子彦刘惠民唐纲岭樊美娟崔华鹏赵乐张晓兵蔡君兰王冰
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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