Immunofluorescence chromatography detection card and method for simultaneous detection of florfenicol and trimethoprim in poultry eggs

A technology of florfenicol and trimethoprim, which is applied in the field of biological detection, can solve the problems of only one of them being detected, high requirements for samples to be tested, and threats to public health, so as to improve sensitivity and labeling efficiency , Reduce the effect of steric hindrance effect

Active Publication Date: 2021-07-02
广州敏捷生物技术有限公司
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  • 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 simultaneous detection of florfenicol and trimethoprim in poultry eggs
  • Immunofluorescence chromatography detection card and method for simultaneous detection of florfenicol and trimethoprim in poultry eggs
  • Immunofluorescence chromatography detection card and method for simultaneous detection of florfenicol and trimethoprim in poultry eggs

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Experimental program
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Effect test

Embodiment 1

[0066] In this embodiment, the immunofluorescent chromatography detecting card in which the fluorophenicon, the methoxymethridin content is synchronized, including the bottom plate, and the bottom plate, and the bottom plate is connected to the left to right, and the water absorbing paper pad, the reaction film, the combined pad, The sample pad, the reaction film was prepared with fluoropheniconi detection line T1, the methoxybenzil detection line T2, and the heat row of three lines, the binding pad was covered with fluorine tissue detection microspheres. Methylene anti-chicken IGY polyclonal antibody labeling microspheres;

[0067] Fluornisi detection microspheres are long-chain biotinoflu fluoropheniconi monoclonal antibodies and streptavidin fluorescent microspheres with streptavidin fluorescent microspheres by connecting polyethylene glycol long-chain excess arms (-DPEG24-). Streptaviarin is combined;

[0068] Methoxybenzeridine detection microspheres are ultra-chain biotin-li...

Embodiment 2

[0106] This embodiment provides a method of detecting a fluorine-based, methoxyenin content synchronously detecting a fluorine content in an egg, and specifically comprises the steps of:

[0107] (1) Pre-treatment of the sample first, the specific steps are: Take 2G to detect homogeneous eggs that do not contain fluorine-containing and methoxybenzil as a negative reference, 3 ml of acetic acid per tube. Ester (containing 1 wt% trichloroacetic acid), manual shock 30s, 4000 rpm, centrifugation for 1 min. Take 1 mL of the supernatant to 15 ml of centrifuge tube, 5 ml of n-hexane, 0.5 ml of reconstitution, and the lower layer water phase to be tested;

Embodiment 3

[0110] Preparation of the standard

[0111] Series standard preparation: Take 16 μL of fluorine tissue standard (10 mg / kg) and 16 μL of methoxymethridine standard (10 mg / kg) to add to 968 μl of methanol solution, mixed with mixed standard 1 (fluorbennette It is 160 μg / kg, methoxybazine is 160 μg / kg), and then doubles 5 times to mix the standard 2, mixed standard 3, mixed standard 4, mixed standard 5, mixed standard 6; take 7 15 ml of centrifuge tube, 1 ml of white samples without fluorine-free and methoxy, and add 50 μl of mixed standard 1-6 to 6 tubes, and 50 μl of methanol is added to mix.

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

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

[0114] The 3 ml extract was added, and the hand was shaken for 30s, 4000 rpm, centrifuged for 2 ...

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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; it aims to provide a detection method that is simple and time-saving in sample pretreatment operation, and does not need to be dried. An immunofluorescence chromatography detection card and method for simultaneous detection of florfenicol and trimethoprim content in poultry eggs with high sensitivity, suitable for on-site instant detection, and its technical points include a bottom plate, the bottom plate is from left to The right side is connected with an absorbent paper pad, a reaction membrane, a binding pad, and a sample pad in turn. The reaction membrane is marked with three lines: the florfenicol detection line T1, the trimethoprim detection line T2 and the quality control C line. The binding pad of the invention is coated with florfenicol detection microspheres, trimethoprim detection microspheres, and goat anti-chicken IgY polyclonal antibody-labeled microspheres; it belongs to the technical field of biological detection.

Description

Technical field [0001] The present invention discloses a mapping egg detection method, and in particular, the present invention discloses a synchronous detection of an immunofluorescent chromatographic detection card and method of forming a fluorine-benny content in an avian egg, belonging to biological detection technology. field. Background technique [0002] Fluorine Niki is a new type of chloramphenicol-spectrum antibiotics, which is significantly advantageous in safety and effectiveness, and is used in a large number of livestock and poultry aquaculture. Methylene Tammyxylphenamine methoxymethrimethyltrimethyltrimethyltrimethyltrimethyltrimethylene, which belongs to the broad-spectrum antibacterial synergist, which is not strong, but combined with sulfonamides and some antibiotics has significant synergies, in livestock and poultry A wide range of industries. [0003] However, the large number of fluorbennette and methoxymetidine has a high detection rate in livestock and po...

Claims

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Application Information

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Patent Type & Authority Patents(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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