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Immune colloidal gold detecting card of diclazuril and preparation method of immune colloidal gold detecting card

A technology of diclazuril and colloidal gold, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complicated operation, expensive equipment, difficult to promote, etc., and achieve the effect of accurate results and simple methods

Inactive Publication Date: 2016-05-11
NANJING YITE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The detection methods of diclazuril in the prior art are mainly high-performance liquid chromatography and other detection methods, but the defects of these technologies are obvious: expensive equipment, complicated operation, difficult to popularize

Method used

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  • Immune colloidal gold detecting card of diclazuril and preparation method of immune colloidal gold detecting card
  • Immune colloidal gold detecting card of diclazuril and preparation method of immune colloidal gold detecting card
  • Immune colloidal gold detecting card of diclazuril and preparation method of immune colloidal gold detecting card

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Experimental program
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Effect test

Embodiment 1

[0021] 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 is coated with diclazuril-BSA and goat anti-mouse IgG; 11 is a water-absorbent pad, made of water-absorbent materials such as filter paper.

[0022] 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.

[0023] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber sheet 9 . Goat anti-mouse IgG quality control line 3 and diclazuril-BSA detection line 2 are set on the nitrocellulose membrane 10 .

[0024] At the other end of the PVC rubber sheet 9, a water-absorbing pad 11 is adhered. One end of the nitrocellulose me...

Embodiment 2

[0026] The preparation of diclazuril colloidal gold detection card is realized by the following steps:

[0027] 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;

[0028] Antibody pretreatment: centrifuge the diclazuril 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 with 0.01mol / LPBS / ml, pass through a 0.22μm filter membrane;

[0029] Preparation of colloidal gold markers: get 40 mL of colloidal gold solution in step (1), adj...

Embodiment 3

[0035] Diclazuril Minimum Detection Limit Test

[0036] Accurately weigh 0.500g diclazuril antigen, dissolve it in ultrapure water and dilute to 100mL, mark it as A 1 solution, take A 1Add 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 3 Put 1mL of the solution in 4 test tubes, and label them as 1~4 respectively. Add 9mL, 4mL, 1mL, and 0mL of ultrapure water to the test tubes respectively, and the concentrations of diclazuril antigen solutions are: 0.5ng / mL, 1ng / 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 diclazuril test card, and observe the results 5 minutes after adding the sample. The color developmen...

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Abstract

The invention discloses an immune colloidal gold detecting card of diclazuril and a preparation method of the immune colloidal gold detecting card, and relates to the technical field of animal source food veterinary drug residue detection. A test strip in a shell of the detecting card is composed of a PVC glue plate, a sample cushion, a colloidal gold conjugate pad, a coating film and a water absorbing cushion, wherein a colloidal gold film is a glass cellulose membrane containing diclazuril monoclonal antibodies, the coating film is a nitrocellulose film and is provided with a line T and a line C, the line T is wrapped by diclazuril protein conjugates, and the line C is wrapped by goat-anti-mouse IgG antibodies. The detecting card can be effectively used for rapidly detecting diclazuril and is convenient to use, rapid and accurate in result.

Description

technical field [0001] The invention relates to the technical field of detection of veterinary drug residues in cereal foods, in particular to an immune colloidal gold detection card for diclazuril and a preparation method thereof. Background technique [0002] Diclazuril is a triazine benzyl nitrile compound, off-white or light yellow powder, slightly soluble in dimethylformamide, slightly soluble in tetrahydrofuran, almost insoluble in water and ethanol. Diclazuril is a broad-spectrum coccidial anthelmintic with high efficiency and low toxicity. It has a good control effect on various coccidia such as chickens, turkeys, ducks, geese, peacocks, quails, rabbits, etc. Add 1g per ton of feed , can effectively control the outbreak of coccidiosis. Chicken coccidia are common parasites in the poultry industry, causing serious economic losses to the poultry industry. At present, the main means of preventing coccidiosis in chickens is to use anticoccidiosis drugs for chemical con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 洪霞杜霞江振飞
Owner NANJING YITE BIOTECH
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