Florfenicol on-site test paper, and preparation and use methods thereof

A florfenicol, on-site detection technology, applied in the field of immunology, can solve problems such as weight reduction

Active Publication Date: 2015-12-09
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Guo Guifang and others believe that FF injection has no obvious acute toxicity and subchronic toxicity, some studies have shown that high doses of FF can reduce the weight of thymus and spleen in mice, and foreign studies have also shown that FF can inhibit the humoral immunity and cellular immunity of mice. Immunity, the conclusions drawn by Guo Guifang confirm each other, indicating that FF has strong immunotoxicity

Method used

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  • Florfenicol on-site test paper, and preparation and use methods thereof
  • Florfenicol on-site test paper, and preparation and use methods thereof
  • Florfenicol on-site test paper, and preparation and use methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The synthesis of florfenicol amine: According to the molar ratio of 1:1, weigh FF10.74, add 2.56g of sodium hydroxide to 15ml of distilled water to dissolve, heat and stir until completely dissolved, at this time the solution is wine red and clear, then add 6.0 g of sodium chloride, stirred until completely dissolved, after cooling, use a thin-layer chromatography plate to chromatograph the mixed solution, fumigate with iodine for color development, and measure the Rf (the ratio of the distance from the origin to the center of the spot to the distance from the origin to the solvent front) value Unlike FFC, it is proven that FFA is generated. Leave the solution at room temperature overnight, filter with filter paper, and wash with distilled water until the effluent is colorless and dried FFA hapten. The synthetic route is as follows:

[0026]

Embodiment 2

[0028]Preparation of complete antigen: Weigh 25mg florfenicol amine and dissolve it in 2mL DMF; weigh 66mg BSA and dissolve it in 10ml pH7.4 PBS buffer; add florfenicol amine dropwise to the above BSA solution, at 4℃ After stirring, 100 μL of glutaraldehyde was added dropwise to the reaction liquid, and the reaction was stirred slowly for 24 h. The synthesized product was dialyzed with PBS for 72 hours, and the solution was changed every 8 hours to obtain the immunogen of florfenicol amine-bovine serum albumin (FFA-BSA), and the same method was used to prepare florfenicol amine-ovalbumin (FFA-OVA) ) is coated with the original, and the synthetic route is as follows:

[0029]

Embodiment 3

[0031] Preparation of anti-florfenicol amine monoclonal antibody: the hybridoma cells secreting anti-florfenicol amine monoclonal antibody of the present invention, its preparation and selection method are as follows:

[0032] (1) Preparation of Florfenicol amine (FFA) hapten by alkaline hydrolysis

[0033] (2) Synthesize the complete antigen of florfenicol amine with the glutaraldehyde method, including the immunogen FFA-BSA, and the former FFA-OVA of the coating;

[0034] (3) Immunizing Balb / c mice with the immunogen FFA-BSA;

[0035] (4) fusion of splenocytes and myeloma cells of immunized mice to obtain 120 strains of hybridoma cells;

[0036] (5) Screening positive hybridoma cell lines by indirect ELISA method coated with original FFA-OVA coated plates;

[0037] (6) Four of the positive hybridoma cells were cloned by the limiting dilution method;

[0038] (7) The antibody prepared by the secondary cloned cell line of the anti-florfenicol amine monoclonal antibody 2D6-1...

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Abstract

The invention particularly relates to florfenicol on-site test paper, and preparation and use methods thereof and belongs to the field of immunology. The test paper includes anti-florfenicol amine secreted from hybridoma FFA-C which is assigned the accession number CCTCC-C201575, a monoclonal antibody C marked by colloidal gold and a carrier plate having adhesive sticker, wherein a sample adding end water absorption layer and a hand holding end water absorption layer are formed at two ends of the carrier plate respectively. A test layer is arranged on the middle of the carrier plate. A glass fiber layer block, on which the monoclonal antibody C marked by colloidal gold is supported, is arranged at the boundary between the test layer and the sample adding end water absorption layer. One end of the glass fiber layer block is arranged under the sample adding end water absorption layer and the other end of the glass fiber layer block is arranged above the test layer. A test line and a quality control line are formed on the test layer extended from the glass fiber layer block, wherein the test line and the quality control line are respectively coated by an anti-florfenicol amine polyclonal antibody and sheep-anti mouse IgG. The test paper quickly, simply and accurately detects the florfenicol and the metabolite, florfenicol amine, thereof on site, and can be used for crude screening test of drug residue of the florfenicol.

Description

technical field [0001] The invention relates to the improvement of antibiotic detection technology, in particular to on-site detection test strips and their preparation and use methods, belonging to the field of immunology. Background technique [0002] Florfenicol (Florfenico, FF) Chinese name: fluprofen, fluthiamphenicol, chemical name: d(+)-threo-1-p-methylphenyl-2-dichloroacetamido-3-fluoro Propanol is a broad-spectrum antibacterial drug of chloramphenicol for animals developed by Schering-Plough of the United States in the late 1980s. It is the third generation of chloramphenicol antibiotics. Listed in Japan, as a substitute for the new chloramphenicol, it mainly inhibits the binding of 50s subunits to inhibit the activity of bacterial peptidyl aminotransferase. It is a new type of anti-Gram-positive bacteria, Gram-negative bacteria and thiamphenicol-resistant bacteria A broad-spectrum antibiotic that can be used to treat bacterial diseases in pigs, cattle, poultry and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/54306G01N33/558
Inventor 王晓洁
Owner LUDONG UNIVERSITY
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