Method for detecting Ennoxacin and its special enzyme-linked immune reagent kit

An enzyme-linked immunosorbent reagent, enrofloxacin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome processes, unsuitable for on-site monitoring and screening of a large number of samples, complex instruments and equipment, etc., and achieve convenient use , high efficiency and high sensitivity

Inactive Publication Date: 2010-04-21
CHINA AGRI UNIV
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  • Abstract
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Problems solved by technology

[0003] 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 equipment and cumbersome processes

Method used

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  • Method for detecting Ennoxacin and its special enzyme-linked immune reagent kit
  • Method for detecting Ennoxacin and its special enzyme-linked immune reagent kit
  • Method for detecting Ennoxacin and its special enzyme-linked immune reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Preparation and detection method of ELISA kit with anti-antibody as coating source

[0032] ELISA kits with anti-antibody conjugates as the coating source include:

[0033] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0034] (2) Alkaline phosphatase-labeled enrofloxacin hapten working solution (alkaline phosphatase-labeled enrofloxacin hapten diluted with an aqueous solution containing 0.05% glycerol), 8ml / bottle, 1 bottle.

[0035](3) Enrofloxacin standard solution: 6 bottles of enrofloxacin series standard solutions, 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L, 3ml / bottle. The enrofloxacin drug diluent used is a phosphate buffer solution containing 0.1-0.5% N'N-dimethylformamide (DMF).

[0036] (4) The chromogenic solution is 4-nitrophenol phosphate buffer solution, 8ml / bottle, 1 bottle.

[0037] (5) Enrofloxacin mouse monoclonal antibody working solution: Dilute the monoclonal antibody secreted by enrofloxacin monoclonal...

Embodiment 2

[0071] Example 2, the precision, accuracy and shelf life experiments of the enrofloxacin ELISA kit

[0072] 1. Standard solution precision test

[0073] Get three batches of 02, 06, and 10 from the ELISA kit prepared in Example 1, get 10 kits from each of the three batches, and extract 20 microwells from each microplate, measure 4.0 Calculate the coefficient of variation from the absorbance value (OD value) of the μg / L standard solution. This method was used to examine the intra-panel precision. The measurement results are shown in Table 1. The results showed that, for the three batches of kits, the measurement range of the standard precision was between 4.80% and 9.11%, so the determined precision range was all less than 25%.

[0074] Table 1 Standard repeatability test (CV%)

[0075]

[0076] 2. Sample repeatability test

[0077]Each sample was added with enrofloxacin standard at a concentration of 10 μg / kg, and three different batches of kits prepared in Example 1 w...

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Abstract

The present invention discloses a method for detecting enoxacin and its special-purpose ELIA kit. Said kit includes enoxacin specific antibody, coating source, enzyme lablling plate and enzyme label,enoxacin standard sample solution, substrate display solution, stop buffer and concentration double solution. The described coating source is enoxacin antiantibody, the described enzyme label is enzymelabeled enoxacin antigen.

Description

technical field [0001] The invention relates to a method for detecting enrofloxacin and a special ELISA kit. Background technique [0002] Enrofloxacin (ENR) is a fluoroquinone 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-β-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 will pose a threat and affect human health. European and American countri...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠何方洋万宇平史为民丁双阳冯才伟江海洋吴小平
Owner CHINA AGRI UNIV
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