Zilpaterol immune colloidal gold detection card and preparation method thereof

A technology of colloidal gold and detection card, which is applied in the direction of measuring device, biological test, material inspection product, etc., to achieve the effect of simple method and accurate result.

Inactive Publication Date: 2014-05-21
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above situation, the purpose of the present invention is to provide a zilpaterol immune colloidal gold detection card and its preparation method ...

Method used

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  • Zilpaterol immune colloidal gold detection card and preparation method thereof
  • Zilpaterol immune colloidal gold detection card and preparation method thereof
  • Zilpaterol immune colloidal gold detection card and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0022] figure 1 It is the structural diagram of the immune colloidal gold detection card of zilpaterol of the present invention, figure 2 It is the cross-sectional structure diagram of the test paper strip in the immune colloidal gold detection card of zilpaterol of the present invention: among the figure 9 is a PVC glue board; 7 is a sample pad; 8 is a colloidal gold bonding pad, and the colloidal gold bonding pad is covered with Colloidal gold marker of monoclonal antibody; 10 is the coated membrane, that is, nitrocellulose membrane, which is coated with zilpaterol-BSA and goat anti-mouse IgG; 11 is the water-absorbent pad, made of water-absorbent material Made of filter paper.

[0023] On one end (sample end) of the PVC rubber plate 9, the sample pad 7 and the colloidal gold bonding pad 8 are adhered, and the sample pad 7 and the colloidal gold bonding pad 8 are in a side-by-side structure.

[0024] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber ...

Embodiment 2

[0027] The preparation of zilpaterol colloidal gold detection card is realized by the following steps:

[0028] (1) Preparation of colloidal gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 (0) 5 mL, prepared into 500 mL 0.01% chloroauric acid aqueous solution, heated and boiled, then added 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Solution 5-7 mL, continue to stir and heat, when the color of the solution turns completely transparent purple red, stop heating after maintaining for 5 min, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;

[0029] (2) Antibody pretreatment: centrifuge the zilpaterol antibody to be labeled at 1000 r / min, 4°C for 20 min, take the supernatant, and dilute it to 1 mg / mL with 0.01 mol / L PBS; or use Dilute 0.01 mol / L PBS to 1 mg / ml, pass through a 0.22 μm filter membrane;

[0030] (3) Preparation of colloidal gold markers: ta...

Embodiment 3

[0036] Zipaterol minimum detection limit test

[0037] Accurately weigh 0.500 g zilpaterol standard substance, dissolve it in ultrapure water and make up to 100 mL, marked as A 1 solution, take A 1 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 2 solution, take A 2 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 3 solution, the concentration of A3 solution is 5 ng / mL (5ppb). take A 3 Put 1 mL of the solution in 4 test tubes, and label them 1~4 respectively, add 9 mL, 4 mL, 1 mL, and 0 mL of ultrapure water to the test tubes respectively, and prepare the concentrations of zilpaterol standard solutions as follows: 0.5 ng / mL, 1 ng / mL, 2.5 ng / mL, 5 ng / mL; take another empty test tube, add 5 mL of ultrapure water, and mark it as No. 5 test tube. Take the liquid in the above test tubes 1-5 as samples and drop them on the zilpaterol test card, and observe the results 5 minutes after adding ...

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Abstract

The invention discloses a zilpaterol immune colloidal gold detection card and a preparation method thereof, and relates to the beta-epinephrine receptor agonist detection technical field. A test strip in an outer shell of the detection card is composed of a PVC rubber plate, a sample pad, a colloidal gold combined pad, a coating film and a water-absorbing pad; a colloidal gold film is a glass cellulose film containing a zilpaterol monoclonal antibody, and the coating film is a nitrocellulose film; a T line and a C line are arranged on the nitrocellulose film, the T line is coated with a zilpaterol-protein conjugate, and the C line is coated with a sheep anti-mouse IgG antibody. The detection card is effectively used for rapid detection of zilpaterol, is convenient and fast, and has accurate results.

Description

technical field [0001] The invention relates to the technical field of beta-adrenoreceptor agonist detection, in particular to an immune colloidal gold detection card for zilpaterol and a preparation method thereof. Background technique [0002] beta 2 -Adrenergic agonists are a common class of growth promoters that, when used in animal farming, increase feed conversion and increase the ratio of muscle to fat in the tissue. beta 2 -Adrenergic agonists belong to catecholamine compounds, which can be divided into anilines, resorcinols and phenols according to their chemical structures. Zipaterol (Zilpaterol) is a novel beta 2 - Adrenergic agonists whose chemical structure (see formula 1) is completely identical to that of common aniline-adrenergic agonists (clenbuterol and albuterol) and phenolic beta-adrenergic agonists (ractopamine) Differently, it is one-tenth as effective as clenbuterol, but more potent than ractopamine. Zipaterol is permitted as a feed additive for...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/74G01N33/54386G01N33/558
Inventor 杜道林洪霞
Owner JIANGSU WISE SCI & TECH DEV
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