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Immunofluorescence chromatography detection card and method for synchronously detecting contents of florfenicol and trimethoprim in poultry eggs

A technology of florfenicol and trimethoprim is applied to the immunofluorescence chromatography detection card for simultaneous detection of florfenicol and trimethoprim in poultry eggs, and in the field of poultry egg detection, it can solve the problem of samples High requirements, only one of them can be detected, threats to public health, etc.

Active Publication Date: 2021-05-14
广州敏捷生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the large-scale use of florfenicol and trimethoprim has a high detection rate in livestock and poultry products, which seriously threatens public health. Therefore, the national food safety standard has set strict detection limits for these two antibiotics. , it is forbidden to use during the laying period of laying hens
[0004] At present, the detection of florfenicol and trimethoprim in the prior art includes chromatography, enzyme-linked method, immunochromatography, etc. Some of these methods rely heavily on high-end equipment, and some have high requirements for the samples to be tested. Complicated, or only one of them can be detected, seriously reducing work efficiency

Method used

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  • Immunofluorescence chromatography detection card and method for synchronously detecting contents of florfenicol and trimethoprim in poultry eggs
  • Immunofluorescence chromatography detection card and method for synchronously detecting contents of florfenicol and trimethoprim in poultry eggs
  • Immunofluorescence chromatography detection card and method for synchronously detecting contents of florfenicol and trimethoprim in poultry eggs

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Experimental program
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Effect test

Embodiment 1

[0066] The immunofluorescence chromatography detection card for synchronously detecting the contents of florfenicol and trimethoprim in poultry eggs provided by this example includes a bottom plate, on which an absorbent paper pad, a reaction membrane, a binding pad, The sample pad, three lines of florfenicol detection line T1, trimethoprim detection line T2 and quality control line C are drawn on the reaction membrane, and the binding pad is coated with florfenicol detection microspheres, Trimethoprim detection microspheres, goat anti-chicken IgY polyclonal antibody labeled microspheres;

[0067] Florfenicol detection microspheres are made by linking long-chain biotinylated florfenicol monoclonal antibody with polyethylene glycol long chain extension arm (-dPEG24-) and streptavidin fluorescent microspheres through biotin and Made with streptavidin;

[0068] Trimethoprim detection microspheres are made by linking long-chain biotinylated trimethoprim monoclonal antibody with p...

Embodiment 2

[0106] The present embodiment provides a method for synchronous detection of florfenicol and trimethoprim content in poultry eggs, specifically comprising the following steps:

[0107] (1) Pre-treat the sample first. The specific steps are: take 2 g of homogeneous poultry eggs that are free of florfenicol and trimethoprim as a negative control substance, and add 3 ml of ethyl acetate to each tube. Ester (containing 1wt% trichloroacetic acid), shake manually for 30s, centrifuge at 4000rpm for 1min. Take 1ml of supernatant to a 15ml centrifuge tube, add 5ml of n-hexane and 0.5ml of reconstituted solution, and let the lower layer of water phase become the solution to be tested after standing for stratification;

Embodiment 3

[0110] Preparation of Assay Brackets

[0111] Preparation of series standard products: Take 16 μl florfenicol standard product (10 mg / kg) and 16 μl trimethoprim standard product (10 mg / kg) and add them to 968 μl methanol solution, and mix well to obtain mixed standard product 1 (flofenicol 160μg / kg, trimethoprim 160μg / kg), and then doubling dilution 5 times to get mixed standard 2, mixed standard 3, mixed standard 4, mixed standard 5, mixed standard 6; take 7 15ml centrifuge tube, add 2ml of blank sample without florfenicol and trimethoprim to each tube, take 50μl mixed standard 1-6 and add it to 6 tubes, add 50μl methanol to the 7th tube, and mix well.

[0112] The concentration gradient of florfenicol is: 4μg / kg, 2μg / kg, 1μg / kg, 0.5μg / kg, 0.25μg / kg, 0.125μg / kg, 0μg / kg;

[0113] The concentration gradient of trimethoprim is: 4μg / kg, 2μg / kg, 1μg / kg, 0.5μg / kg, 0.25μg / kg, 0.125μg / kg, 0μg / kg;

[0114] Add 3ml of extract solution to each tube, shake manually for 30s, centrifuge ...

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Abstract

The invention discloses an immunofluorescence chromatography detection card and method for synchronously detecting the contents of florfenicol and trimethoprim in poultry eggs, and aims to provide an immunofluorescence chromatography detection card and method which are simple and time-saving in sample pretreatment operation, free of blow-drying, high in detection method sensitivity, suitable for on-site instant detection and capable of synchronously detecting florfenicol and trimethoprim in poultry eggs. The card is characterized by comprising a bottom plate, an absorbent paper pad, a reaction film, a combination pad and a sample pad are sequentially connected to the bottom plate from left to right, a florfenicol detection line T1, a trimethoprim detection line T2 and a quality control line C are marked on the reaction film, and florfenicol detection microspheres, trimethoprim detection microspheres and goat anti-chicken IgY polyclonal antibody labeled microspheres are coated on the combination pad; the invention belongs to the technical field of biological detection.

Description

technical field [0001] The invention discloses a poultry egg detection method. Specifically, the invention discloses an immunofluorescence chromatography detection card and method for synchronously detecting the contents of florfenicol and trimethoprim in poultry eggs, belonging to biological detection technology field. Background technique [0002] Florfenicol is a new type of broad-spectrum chloramphenicol antibiotic with obvious advantages in safety and effectiveness, and is widely used in livestock and poultry farming. The full name of trimethoprim is trimethoprim, which is a broad-spectrum antibacterial synergist. Its own bactericidal ability is not strong, but it has a significant synergistic effect when used in combination with sulfa drugs and some antibiotics. It is used in livestock and poultry breeding. widely used in industry. [0003] However, the large-scale use of florfenicol and trimethoprim has a high detection rate in livestock and poultry products, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/58G01N33/533
CPCG01N33/533G01N33/558G01N33/577G01N33/582G01N33/585G01N33/9446
Inventor 汤永平刘卫林志欣羊晓珊梁展鹏高成李立
Owner 广州敏捷生物技术有限公司
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