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Immune chromatography test paper for quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and preparation method thereof

A technology of fluorescent nanoparticles and immunochromatographic test paper, which is applied to measuring devices, analytical materials, instruments, etc., to achieve the effects of eliminating interference, displaying images, and improving sensitivity

Active Publication Date: 2012-11-28
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After retrieval, UCP immunochromatographic test paper used to detect pesticide and veterinary drug residues in food has not been reported. Although colloidal gold test paper is widely used in this area, it cannot be quantitatively detected. As a more sensitive, stable, flexible and safe The UPT-LF research is a powerful supplement to the colloidal gold immunoassay technology, and it is urgent to strengthen the research and discussion in this aspect

Method used

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  • Immune chromatography test paper for quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and preparation method thereof
  • Immune chromatography test paper for quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and preparation method thereof
  • Immune chromatography test paper for quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and preparation method thereof

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Experimental program
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preparation example Construction

[0051] 1. Preparation of CL monoclonal antibody or polyclonal antibody

[0052] Preparation of monoclonal antibody: including preparation of CL immunogen, immunization of animals (mice), cell fusion, screening of monoclonal antibody, preparation of monoclonal antibody; preparation of polyclonal antibody: preparation of CL immunogen, immunization of animals (rabbit), screening of polyclonal antibodies, preparation of polyclonal antibodies.

[0053] (1) Preparation of CL artificial antigen:

[0054] Dissolve 100 mg CL standard in 10mL, 1mol / 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 CL solution dropwise into PBS (pH7.4) dissolved in 300 mg BSA with a dropper, ...

Embodiment 1

[0087] Embodiment one: see figure 1 with figure 2 . In the figure, 1 is the support layer, made of plastic sheet strips, 2 is the adsorption fiber layer, made of glass fiber cotton, fluorescent antibody fiber layer 3 is adsorbed with fluorescent antibody glass fiber cotton of anti-CL monoclonal antibody, cellulose membrane Layer 4 is made of nitrocellulose membrane, and the water-absorbing material layer 5 at the handle end is made of water-absorbing filter paper, and the layers of absorbent fiber layer 2, fluorescent antibody fiber layer 3, cellulose film layer 4, and water-absorbing material layer 5 are pasted in sequence from right to left Fixed on the support layer 1, the fibers cross and penetrate each other at the junction. On the cellulose membrane layer 4, there is a stealth detection blot 6, which is made with CL-coupled bovine serum albumin solution (BSA); the stealth control blot 7 is blotted on the cellulose membrane with a goat anti-mouse IgG antibody solution...

Embodiment 2

[0093] Embodiment 2: The structure of the test strip is basically the same as that of Embodiment 1, the difference is that the fluorescent antibody fiber layer 3 is adsorbed with anti-CL polyclonal antibodies, the adsorption fiber layer 2 is made of nylon membrane, and the cellulose membrane layer 4 is made of The pure cellulose film, the invisible detection imprinted band and the invisible contrast imprinted band are all "ten", and 8-2 is the blue protective film at the handle end covered on the water-absorbing material layer 5.

[0094] Detection of milk samples: dilute the milk samples with physiological saline to make a sample suspension of 1:2~5.

[0095] Result judgment: (a) Positive If there is a red blot on the cellulose membrane " ten ", indicating that the test result is positive, indicating that CL is contained in the sample to be tested; (b) negative if there are two red blots on the cellulose membrane" ten " means that the test result is negative, indicating th...

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Abstract

The invention discloses an immune chromatography test paper strip for the quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and a preparation method thereof. The test paper strip comprises a supporting layer, an adsorption layer, and a protection layer, wherein the adsorption layer comprises an adsorption fibrous layer, a fluorescent antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end, the cellulose film is provided with detection blots printed by using a CL coupling carrier protein solution and control blots printed by using goat anti-mouse IgG antibodies, and the fluorescent antibody is an NaYF4:Yb:Er nanoparticle labelled CL monoclonal antibody or polyclonal antibody. According to the invention, the application of immune chromatography based on the up-conversion fluorescent nanoparticle label in the quantitative determination of CL residues is realized, so that the detection of the CL residues has no background interference; and the test paper strip has the advantages of strong specificity, high sensitivity, simple, and accurate detection, low cost, wide application scope, and easiness in popularization and application.

Description

technical field [0001] The invention relates to an immunochromatographic test strip, in particular to an immunochromatographic test strip for quantitative detection of clenbuterol residues and a preparation method thereof, using more than one converted fluorescent material as a biomarker. Background technique [0002] Clenbuterol (Clenbuterol, CL), commonly known as "Clenbuterol", Clenbuterol hydrochloride is its hydrochloride, which can promote muscle development and fat decomposition. Clenbuterol can improve the carcass lean meat of meat animals Rate. Because consumers have a special preference for lean meat, pigs with a high yield of lean meat can be sold at a good price, and the addition of "clenbuterol" has brought additional benefits to farmers. Therefore, driven by illegal interests, farmers often add "clenbuterol" to animal feed during the breeding process. [0003] But clenbuterol is an optional beta 2 - Adrenergic receptor agonists have great side effects, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/533
Inventor 张改平职爱民宋春美胡骁飞杨继飞赵东
Owner HENAN ACAD OF AGRI SCI
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