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

An immunochromatographic test strip, melamine technology, applied in biological testing, measuring devices, analytical materials, etc., can solve the problem of large-throughput rapid screening and detection not suitable for grass-roots enterprises and institutions, high professional requirements for operators, and matrix effects. Large background interference, etc., to achieve the effect of increasing stability and fluorescence lifetime, increasing reaction sensitivity, and high sensitivity

Active Publication Date: 2018-08-10
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the above methods require advanced detection equipment, the detection costs are expensive, the steps are cumbersome and time-consuming, and the requirements for the professionalism of operators are high, they are not suitable for large-throughput rapid screening detection in grassroots enterprises and institutions.
Colloidal gold immunochromatography has the advantages of fast detection speed, low price, and simple operation. It is currently the main monitoring method for detecting melamine at home and abroad, but there are still some defects, such as poor stability, low sensitivity, and quantitative detection cannot be realized. , The matrix effect is obvious, the background interference is large, and the color is single, it is difficult to realize multiple detection and joint detection

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 detects the composition of the fluorescent microsphere immunochromatography test strip that melamine remains

[0038] The test strip ( figure 1 , figure 2 ) is composed of a bottom plate, a sample binding pad, a nitrocellulose membrane and an absorbent pad;

[0039] 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 start of the sample binding pad is aligned with the start of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom plate;

[0040] A detection area 4 and a quality control area 5 are fixed on the nitrocellulose membrane, the detection area is sprayed with a melamine hapten-carrier protein conjugate (melamine hapten-ovalbumin conjugate), and ...

Embodiment 2

[0042] Example 2 Preparation of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Residual Melamine

[0043] The preparation method of the fluorescent microsphere immunochromatographic test strip for detecting residual melamine mainly comprises the following steps:

[0044]1) Preparation of the sample binding pad: mark the anti-melamine 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;

[0045] 2) Preparation of nitrocellulose membrane: Spray the melamine hapten-carrier protein conjugate on the detection area range on the nitrocellulose membrane to make the detection area; spray the goat anti-mouse anti-antibody onto the nitrocellulose membrane The scope of the quality control area is made into the quality control area;

[0046] 3) Assembly and cutting: Paste the sample binding pad embedded wi...

Embodiment 3

[0088] Example 3 Application of Fluorescent Microsphere Immunochromatography Test Strips to Detect Residual Melamine

[0089] 1. Sample pretreatment

[0090] Place the milk sample to be tested in a water bath at 60-70°C for 5 minutes, take it out, and wait for test.

[0091] 2. Test with test strips

[0092]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 item to be tested through the touch screen , press the "Start Test" button, the fluorescence detector will automatically scan the test paper card; read or print the test results on the display screen of the instrument.

[0093] 3. Analysis of test results

[0094] After the test is completed, the instrument will automatically calculate the concentration of melamine in the milk sample according to the...

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Abstract

The invention discloses a fluorescent microsphere immunochromatographic test strip detecting melamine residue and a preparation method and an application thereof. The test strip comprises a bottom plate, and a sample bonding pad, a nitrocellulose membrane and a water absorbing pad which are successively overlapped and bonded on the bottom plate, wherein a fluorescent microsphere labeled anti-melamine monoclonal antibody is buried on the sample bonding pad, a detection region and a quality control region are fixed on the nitrocellulose membrane, a melamine hapten-carrier protein conjugate is sprayed to the detection region, and a goat anti-mouse antibody is sprayed to the quality control region. The test strip disclosed by the invention is primarily used for detecting the melamine residue and has the advantages of being high in sensitivity, fast to detect, convenient to operate, economical and practical and the like.

Description

technical field [0001] The invention belongs to the field of detection of illegal additives in food safety, and in particular relates to a fluorescent microsphere immunochromatographic test strip for detecting melamine residue in a sample, a preparation method and application thereof. technical background [0002] Melamine is referred to as triamine for short, its chemical name is 1,3,5-triazine-2,4,6-triamine, and its molecular formula is C 3 N 6 h 6 , with a relative molecular mass of 126.12, is a low-toxic and odorless pure white monoclinic prism crystal, widely used in coatings, building materials, papermaking, leather, textiles and other industries. Because the nitrogen content of melamine is as high as 66%, which is 4 times of the average nitrogen content of protein, but the commonly used Kjeldahl nitrogen method cannot distinguish this non-protein nitrogen when measuring the protein content in a sample, so it is used by unscrupulous traders to add In feed and food ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/533
CPCG01N33/533G01N33/68
Inventor 冯才伟何方洋万宇平冯静杨昌松崔廷婷崔彦虎魏力杰
Owner BEIJING KWINBON BIOTECH
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