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Time resolution fluorescence immunoassay kit used for detecting enrofloxacin

A technology of time-resolved fluorescence and enrofloxacin, applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of being unsuitable for on-site monitoring and screening of a large number of samples, cumbersome processes, complex instrument settings, etc., and achieves easy operation , Wide application, strong specific effect

Inactive Publication Date: 2019-05-14
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical methods for detecting enrofloxacin residues mainly include thin layer chromatography (TLC), gas chromatography (GC), high performance liquid chromatography (HPLC), gas-mass spectrometry (GC / MS), liquid-mass spectrometry (HPLC / MS), capillary electrophoresis (CE), etc., are not suitable for on-site monitoring and screening of a large number of samples due to complex instrument settings and cumbersome processes
At present, there are no patents and literature reports on the time-based fluorescent immunoassay for the detection of enrofloxacin

Method used

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Experimental program
Comparison scheme
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Embodiment

[0022] 1. Preparation of immunogen and coating agent

[0023] Synthesis of the immunogen (enrofloxacin-BSA) of the present invention: Accurately weigh 324 mg of enrofloxacin and dissolve it in 2 mL of N,N-dimethylformamide, add γ-aminobutyric acid solution drop by drop under stirring, stir React for 3 hours, and adjust the pH of the reaction solution to about 10. The precipitate was removed by centrifugation. Add the above reaction dropwise to the BSA solution (320mg BSA dissolved in 5mL physiological saline), then add 23mg of N-hydroxysuccinimide (NHS), 45.4mg of N,N-dicyclohexylcarbodiimide (DCC), React overnight at 4°C, remove the precipitate by centrifugation, take the supernatant and dialyze it with phosphate buffered solution (PBS) for 3 days, change the dialysate every 6 hours, freeze-dry the obtained product, and store it at -20°C for later use;

[0024] Synthesis of the coating agent (enrofloxacin-OVA): In the above reaction, after replacing BSA with OVA, the reacti...

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Abstract

The invention discloses a time resolution fluorescence immunoassay kit used for detecting enrofloxacin. The time resolution fluorescence immunoassay kit used for detecting enrofloxacin consists of a porous coated plate, buffer solution, an enrofloxacin standard substance, an enrofloxacin antibody freeze-drying product, a europium-labeled goat-anti-mouse antibody, cleaning solution and enhancementliquid. The detection method for the time resolution fluorescence immunoassay kit used for detecting enrofloxacin comprises the following steps that: (1) preparing immunogen; (2) preparing a coating antigen; (3) preparing a monoclonal antibody; and (4) carrying out sample pretreatment and detection. The kit has the advantages of being short in detection time, high in average yield, simple in sample pretreatment, wide in application, low in detection cost, high in detection specificity, small in within-run and between-run difference, high in sensitivity, and simple and quick in operation, can carry out field operation detection, and is especially suitable for detecting mass samples.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a time-resolved fluorescence immunoassay kit for detecting enrofloxacin. Background technique [0002] Enrofloxacin is a fluoroquinolone antibiotic with the following characteristics: Due to its unique chemical structure, it has become a very important antibiotic in veterinary clinical medicine. Enrofloxacin has broad-spectrum antibacterial activity against Gram-negative bacteria, Gram-positive bacteria and mycoplasma-like pathogenic bacteria. Bacteria, such as anti-p-lactobactin, aminoglycoside, etc. Because of its unique effects, enrofloxacin has been widely used in the prevention and treatment of diseases in animals, fish and shrimp since the 1980s. However, recent studies have found that any animal that has used enrofloxacin has residues in its products within a certain period of time. Because of its neurotoxicity and nephrotoxicity, residues in animal food w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/558G01N33/577
Inventor 杜道林薛永来杜霞
Owner JIANGSU WISE SCI & TECH DEV