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Up-conversion fluorescence immune chromatography test paper for quantitative detection of neomycin and preparation method thereof

A technique of fluorescence immunochromatography and immunochromatography test paper, which is applied in measurement devices, analytical materials, instruments, etc., to achieve the effect of eliminating interference, displaying image and improving sensitivity

Inactive Publication Date: 2013-12-18
河南百奥生物工程有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although colloidal gold test paper is widely used in this area, it cannot achieve quantitative detection. As a more sensitive, stable, flexible and safe UPT-LF research, it is a powerful supplement to colloidal gold immunoassay technology, and it is urgent to strengthen this aspect. research and discussion

Method used

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  • Up-conversion fluorescence immune chromatography test paper for quantitative detection of neomycin and preparation method thereof
  • Up-conversion fluorescence immune chromatography test paper for quantitative detection of neomycin and preparation method thereof
  • Up-conversion fluorescence immune chromatography test paper for quantitative detection of neomycin and preparation method thereof

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preparation example Construction

[0041] Preparation of monoclonal antibodies: including preparation of NEO immunogens, immunization of animals (mice), cell fusion, screening of monoclonal antibodies, preparation of monoclonal antibodies; preparation of polyclonal antibodies: preparation of NEO immunogens, immunization of animals (rabbit), screening of polyclonal antibodies, preparation of polyclonal antibodies.

[0042] (1) Preparation of NEO artificial antigen:

[0043] Dissolve 100 mg NEO standard in 10 mL, 1 mol / L hydrochloric acid, slowly add 30% NaNO 2 The solution was detected with starch KI test paper, and the dropwise addition was stopped when the KI test paper turned brownish purple, and the reaction was stirred for 45 min, and 5% ammonium sulfamate solution was added dropwise, and the reaction end point was detected with starch KI test paper. Slowly add the azated NEO solution dropwise into 300 mg BSA-dissolved PBS (pH 7.4) with a dropper, adjust the pH to 9.0 with 1 mol / L NaOH solution, and react ...

Embodiment 1

[0076] Embodiment one: see figure 1 with figure 2 . In the figure, the support layer 1 is made of plastic sheet strips, the adsorption fiber layer 2 is made of glass fiber cotton, the fluorescent antibody fiber layer 3 is adsorbed with fluorescent antibody glass fiber cotton of anti-NEO monoclonal antibody, and the cellulose membrane layer 4 is made of nitric acid The cellulose membrane, the water-absorbing material layer 5 at the handle end is made of water-absorbing filter paper, and the layers of the absorbent fiber layer 2, the fluorescent antibody fiber layer 3, the cellulose film layer 4, and the water-absorbing material layer 5 are pasted and fixed on the support layer in sequence from right to left 1, the fibers at the junction between each other interpenetrate each other. On the cellulose membrane layer 4, there is a stealth detection blot 6, which is made with NEO-coupled bovine serum albumin solution (BSA); the stealth control blot 7 is made by blotting a goat a...

Embodiment 2

[0082] Embodiment 2: Immunochromatography test paper is basically the same as Embodiment 1, the difference is that: the fluorescent antibody fiber layer is adsorbed with anti-NEO polyclonal antibody, the adsorption fiber layer is made of nylon membrane, and the cellulose membrane layer is made of pure cellulose The film, the invisible test blot and the invisible control blot are all "ten", and the handle end protective film covering the water-absorbing material layer is blue. Detection of milk samples: dilute the milk samples with physiological saline to make a sample suspension of 1:2~5. Result judgment, operation method are the same as embodiment one.

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Abstract

The invention discloses an up-conversion fluorescence immune chromatography test paper for the quantitative detection of neomycin and a preparation method thereof. The up-conversion fluorescence test paper comprises a support layer, an adsorption layer and a protection layer; the absorbing layer comprises an adsorption fiber layer, a fluorescent antibody fiber layer, a cellulose membrane layer and a water adsorption material layer at a handle hand; the cellulose membrane layer is provided with detection blotting printed by a carrier protein solution coupled with NEO and contrast blotting printed by a goat anti-mouse IgG; the fluorescent antibody adopts an NEO monoclonal antibody or polyclonal antibody marked by NaYF4:Yb:Er nanoparticles. Through the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin, the application of the immune chromatography marked by up-conversion fluorescence nanometer materials in the quantitative detection of NEO residual is realized, so that the detection of the NEO residual is not subjected to background interference; the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin is strong in specificity, high in sensitivity, simple, intuitional and accurate in detection, low in cost, wide in range of application and easy to popularize and apply.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to an up-conversion fluorescence immunochromatographic test paper for quantitative detection of neomycin and a preparation method thereof. Background technique [0002] Neomycin (NEOenbuterol, NEO), neomycin belongs to the glucosamine class of antibiotics, with a broad antibacterial spectrum, commonly used to inhibit bacterial gastrointestinal tract infection in livestock and poultry. With the widespread application of neomycin in the breeding industry at home and abroad, the problem of its residue in food of animal origin has become increasingly prominent. Long-term consumption of animal products with excessive neomycin residues can produce chronic cumulative toxic effects on the body. Announcement No. 235 issued by the Ministry of Agriculture of my country stipulates that the maximum residue limit (MRL) of neomycin in the muscle, liver and fat of cattle, pigs, sheep, chicke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/533
Inventor 张改平职爱民王方雨宋春美胡骁飞王栋
Owner 河南百奥生物工程有限公司
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