Immunocolloidal-gold detection card for kitasamycin and preparation method thereof

A technology of kitasamycin and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of urgent residue detection and hidden dangers of human health, and achieve the effect of simple method and accurate results

Inactive Publication Date: 2017-02-15
ZHENJIANG YITE BIOTECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its residues in animal-derived foods have brought great hidden dangers to human health.
Therefore, residue detection is urgent

Method used

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  • Immunocolloidal-gold detection card for kitasamycin and preparation method thereof
  • Immunocolloidal-gold detection card for kitasamycin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] figure 1 It is the structural diagram of the immune colloidal gold detection card of kitasarmycin of the present invention, figure 2 It is a sectional structure diagram of the test strip in the immune colloidal gold detection card of kitasamycin of the present invention: among the figure 9 is a PVC rubber sheet; 7 is a sample pad; 8 is a colloidal gold binding pad, which is coated with Monoclonal anti-soil colloidal gold marker; 10 is a coated membrane, that is, a nitrocellulose membrane, which is coated with kitasamycin-BSA and goat anti-mouse IgG; 11 is a water-absorbent pad, made of water-absorbent materials such as filter paper become.

[0019] On the other 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.

[0020] A nitrocellulose membrane 10 is adhered in the middle of the PVC colloid 9, and a goat anti-mouse Ig...

example 2

[0023] The preparation of kitasamycin colloidal gold detection card is realized by the following steps:

[0024] (1) Preparation of gelatinous gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 5 mL of 1% chloroauric acid to prepare 500 mL of 0.01% chloroauric acid aqueous solution, heat to boil and then continue stirring 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;

[0025] (2) Antibody pretreatment: centrifuge the kitamycin antibody to be labeled at 1000 r / min, 4°C for 20 minutes, take the supernatant, and dilute it to 1 mg / mL with 0.01mol / L PBS; or use 0.01mol / L Dilute L PBS to 1mg / mL, pass through a 0.22 um filter membrane;

[0026] (3) Preparation of colloidal gold markers: tak...

Embodiment 3

[0032] Kitamycin minimum detection limit test

[0033] Accurately weigh 0.500g of kitasarmycin standard substance, dissolve it in ultrapure water and dilute to 100mL, marked as A 1 solution, take A 1 Add 1 mL of solution to a volumetric flask and add ultrapure water to 1000 mL to form A 2 solution, take A 2 Add 1mL of the solution to a volumetric flask and add ultrapure water to 1000mL to form A 3 solution, then A 3 The concentration of the solution was 5 ng / mL (5 ppb). take A 3 Put 1mL of the solution in 4 test tubes, and label them 1~4 respectively, add 9mL, 4mL, 1ml, 0 ml, ultrapure water to the test tubes respectively, and make A kitasarmycin standard solution, the solubility of which is: 0.5ng / mL.1ng / mL. 2.5ng / mL, 5ng / mL; take another empty test tube, add 5mL of 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 kitasamycin test card, and observe the results 5 minutes after adding the...

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Abstract

The invention provides an immunocolloidal-gold detection card for kitasamycin and a preparation method thereof, belonging to the technical field of detection of antibiotics. A test strip in the shell of the detection card is composed of a PVC rubber sheet, a sample pad, a colloidal-gold binding pad, a coating film and a water-absorbing pad, wherein a colloidal-gold film is a glass cellulose film containing monoclonal antibody against kitasamycin; the coating film is a cellulose nitrate film which is provided with a T line and C line; the T line is coated with a kitasamycin protein conjugate; and the C line is coated with a goat anti-mounse antibody. The immunocolloidal-gold detection card is used for rapid detection of kitasamycin, is convenient and fast and produces accurate results.

Description

technical field [0001] The invention can be effectively used for determining the antibiotic kitasamycin, and the method is simple, convenient, fast and the result is accurate. Background technique [0002] Kitasamycin, also known as leucomycin, is a multi-component macrolide antibiotic produced by Streptomyces kit asatoensis, containing A1, A3, A4, A5, etc. 9 Group A and 4 Group B components. It has strong antibacterial activity against most Gram-positive bacteria and some Gram-negative bacteria and mycoplasma[3], and is used to prevent bacterial infections and mycoplasma diseases in livestock and poultry, as well as as a growth-promoting agent. Kitamycin has a strong bitter taste, which can affect the palatability of animals and stimulate the stomach. Its alkaline characteristics make it degrade in the acidic environment of the stomach (p H ≤ 5.5) and reduce its activity. In order to avoid the above shortcomings, it has been successfully developed Kitamycin microcapsule p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 洪霞刘静
Owner ZHENJIANG YITE BIOTECH DEV CO LTD
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