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ELISA detection method for quantitative determination of cephalosporin antibiotic content in animal-derived food

A technology for detecting animals and detection methods, which is applied in the field of immunoassay to achieve the effects of high accuracy, simple pretreatment and high sensitivity

Inactive Publication Date: 2008-12-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, there are many kinds of cephalosporin antibiotics, and the existing microbial detection methods and instrumental detection methods are difficult to quickly and accurately detect all cephalosporin antibiotics. It is necessary to study a fast and portable multi-residue immunoassay technology

Method used

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  • ELISA detection method for quantitative determination of cephalosporin antibiotic content in animal-derived food
  • ELISA detection method for quantitative determination of cephalosporin antibiotic content in animal-derived food
  • ELISA detection method for quantitative determination of cephalosporin antibiotic content in animal-derived food

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1: The preparation steps of a kit for quantitatively detecting the content of cephalosporin antibiotics in food of animal origin are as follows:

[0033] (1) preparing cephalexin-KLH immunogen;

[0034] (2) Prepare polyclonal antibodies against cephalosporin antibiotics;

[0035] (3) prepare cephalexin-BSA coating former;

[0036] (4) Prepare coated plate;

[0037] (5) Prepare the reagent group.

[0038] Wherein: (1) preparation of cephalexin-KLH immunogen

[0039] Weigh 36.5 mg of cephalexin and dissolve it in 8 mL of 0.01 mol / L, pH 7.4 phosphate buffered saline (PBS), slowly add 40 μL of 25% glutaraldehyde solution under stirring, stir and react at room temperature for 30 min, then add 15 mg of KLH PBS solution was added dropwise, the reaction was stirred at room temperature for 3 h, and then dialyzed in PBS at 4° C. for 72 h. The protein concentration was then determined by the Coomassie brilliant blue method. Store at 4°C for later use.

[0040] (2...

Embodiment 2

[0060] Embodiment 2: The specific application of the present invention is further described through a specific example below.

[0061] Detect the content of cephalosporin antibiotics in milk, taking cephalexin as an example:

[0062] (1) Sample pretreatment: Skim milk can be directly used for detection, while whole milk needs to be centrifuged to remove the fat layer before detection.

[0063] (2) Take out the coated plate, add 100ul cephalexin gradient diluted standard solution and 100ul processed milk sample in parallel to the respective microwells of the coated plate, and then add the cephalexin standard solution and sample Add 100ul of 1:100,000-fold diluted polyclonal antibody to each microwell of the extract, and place at 37°C for 1 hour;

[0064] (3) Wash the plate 3 times with the diluted washing solution, and then add 100ul of color developing solution A and color developing solution B mixed at a ratio of 1:5 to the respective microwells of cephalexin standard soluti...

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Abstract

The invention relates to an enzyme-linked immune detection method for quantitatively detecting the content of spore antibiotic in animal food and belongs to the immune analysis field. The method comprises the steps of adopting the polyclonal antibody of the spore antibiotic with a group-specific antibody as a detection reagent; preparing an enzyme-linked immune detection kit; by using cephalexin as a standard, measurating the concentration of different spore class antibiotics and the relative value of the cephalexin standard through an experiment; and providing the measurated correction factors. The multi-residue detection of the enzyme-linked immune detection kit is expected to be realized. The method has the advantages of simple pretreatment, high sensitivity, high accuracy, etc.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay method for quantitatively detecting the content of cephalosporin antibiotics in animal source food, belonging to the field of immune analysis. Background technique [0002] Cephalosporins are β-lactam antibiotics. In animal husbandry, β-lactam antibiotics are widely used to control mastitis in dairy cows, and treat animal urinary tract, gastrointestinal tract and respiratory tract infections. However, due to its improper use or non-compliance with the withdrawal period regulations, etc., it can cause its residues in animal products, which will bring serious harm to human health, such as allergic reactions, destroying the balance of gastrointestinal flora and enhancing bacteria drug resistance, etc. Therefore, its residue in dairy products has attracted more and more attention at home and abroad. Among them, cephalosporin antibiotics are widely used in the prevention and treatment of human and ani...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/25
Inventor 胥传来谢会玲袁媛赵金山胡拥明
Owner JIANGNAN UNIV
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