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Neomycin immuno-colloidal gold detection card and preparation method thereof

A technology of colloidal gold and neomycin, which is applied in the field of animal source food antibiotic detection, and achieves the effect of simple method and accurate result.

Inactive Publication Date: 2016-04-20
ZHENJIANG XIANCHUANG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above situation, the purpose of the present invention is to provide a neomycin immune colloidal gold detection card and a preparation method thereof in order to overcome the defects in the prior art, which can effectively solve the problem of rapid and easy detection of β-adrenergic receptors. Problems with the body agonist neomycin

Method used

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  • Neomycin immuno-colloidal gold detection card and preparation method thereof
  • Neomycin immuno-colloidal gold detection card and preparation method thereof
  • Neomycin immuno-colloidal gold detection card and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] figure 1 , figure 2 Shown embodiment: among the figure 9 is a PVC rubber sheet; 7 is a sample pad; 8 is a colloidal gold binding pad, and the colloidal gold binding pad is coated with a monoclonal antibody colloidal gold marker; 10 is a coating film, i.e. nitrocellulose Plain membrane, the nitrocellulose membrane coated with neomycin-BSA and goat anti-mouse IgG; 11 is a water-absorbent pad, made of water-absorbent materials such as filter paper.

[0021] On one end (sample end) of the PVC rubber plate 9, the sample pad 7 and the bonding pad 8 are adhered, and the sample pad 7 and the bonding pad 8 are in a side-by-side structure.

[0022] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber sheet 9 . Goat anti-mouse IgG quality control line 3 and neomycin-BSA detection line 2 are set on the nitrocellulose membrane 10 .

[0023] At the other end of the PVC rubber sheet 9, a water-absorbing pad 11 is adhered. One end of the nitrocellulose membrane ...

Embodiment 2

[0025] The preparation of neomycin colloidal gold detection card is realized by the following steps:

[0026] Preparation of colloidal gold solution: Take a 1L Erlenmeyer flask, add 495mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 (0) 5mL, be prepared into 500mL0.01% chloroauric acid aqueous solution, add 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) solution 5-7mL, continue to stir and heat, when the color of the solution turns into transparent purple red completely, stop heating after maintaining 5min, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;

[0027] Antibody pretreatment: Centrifuge the neomycin antibody to be labeled at 1000r / min, 4°C for 20min, take the supernatant, and dilute to 1mg / mL with 0.01mol / LPBS; or dilute to 1mg / mL with 0.01mol / LPBS ml, pass through a 0.22μm filter membrane;

[0028] Preparation of colloidal gold markers: get 40 mL of colloidal gold solution in step (1), adjus...

Embodiment 3

[0034] Neomycin minimum detection limit test

[0035] Accurately weigh 0.500g neomycin antigen, dissolve it in ultrapure water and dilute to 100mL, mark it as A 1 solution, take A 1 Add 1mL of the solution to the volumetric flask and add ultrapure water to 1000mL to form A 2 solution, take A 2 Add 1mL of the solution to the volumetric flask and add ultrapure water to 1000mL to form A 3 solution, the concentration of A3 solution is 5ng / mL (5ppb). take A 3Put 1mL of the solution in 4 test tubes, and label them 1~4 respectively. Add 9mL, 4mL, 1mL, and 0mL of ultrapure water to the test tubes respectively, and make the neomycin antigen solutions with concentrations of 0.5ng / mL and 1ng / mL respectively. mL, 2.5ng / mL, 5ng / mL; take another empty test tube, add 5mL ultrapure water, and mark it as No. 5 test tube. Take the liquid in the above test tubes 1-5 as a sample and add it dropwise to the neomycin test card, and observe the results 5 minutes after adding the sample. The col...

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Abstract

The invention discloses a neomycin immuno-colloidal gold detection card and a preparation method thereof, and relates to the technical field of beta-adrenergic receptor agonist detection. The test strip on the shell of the detection card is composed of a PVC rubber sheet, a sample pad, a colloidal gold combined pad, a coating membrane, and a water absorbing pad; the colloidal gold membrane is a glass cellulose membrane containing a neomycin monoclonal antibody; the coating membrane is a nitrocellulose membrane and is provided with a T line and C line, neomycin protein conjugate is coated in the T line, and goat anti mouse IgG antibody is coated in the C line. The provided detection card is used to detect neomycin in a short time, and has the advantages of convenience, quickness, and precise results.

Description

technical field [0001] The invention relates to the technical field of animal source food antibiotic detection, in particular to an immunocolloid gold detection card for neomycin and a preparation method thereof. Background technique [0002] Neomycin is a broad-spectrum antibiotic commonly used in the treatment of various infectious diseases in animals. It has a strong inhibitory effect on a variety of pathogenic bacteria and is effective on both Gram-positive and Gram-negative bacteria. Neomycin has serious side effects, including nephrotoxicity and inner ear toxicity, and its damage to the inner ear is often irreversible. Therefore, neomycin residues in animal foods pose a huge threat to human health. It is more direct and convenient to detect the content of neomycin in food of animal origin, and it can realize pre-mortem detection. In order to eliminate food safety hazards in time and ensure the quality and safety of food, this test method is specially developed. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/532
Inventor 洪霞杜霞江振飞
Owner ZHENJIANG XIANCHUANG BIOTECH CO LTD
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