Liquid-phase chip method for detecting chloromycetin in aquatic product

A liquid phase chip detection and chloramphenicol technology, applied in the field of bioengineering, can solve the problems of single, time-consuming and high cost of veterinary drug residue detection, and achieve the effects of convenient clinical sampling and analysis, good repeatability and low cost

Inactive Publication Date: 2013-11-13
SHANGHAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It aims to solve the problems of single, time-consuming, and high-cost veterinary drug residue detection, and also lays the foundation for further simultaneous detection of four veterinary drug residue indicators

Method used

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  • Liquid-phase chip method for detecting chloromycetin in aquatic product
  • Liquid-phase chip method for detecting chloromycetin in aquatic product
  • Liquid-phase chip method for detecting chloromycetin in aquatic product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 prepares chloramphenicol artificial antigen

[0048] Chloramphenicol was coupled with ovalbumin to form an artificial antigen.

Embodiment 2

[0049] Preparation and purification of embodiment 2 chloramphenicol monoclonal antibody

[0050] The hapten succinylchloramphenicol containing the shuttle group is coupled to the carrier protein (OVA) by the active ester method, the hapten containing the aromatic amino group is coupled to the carrier protein by the diazotization method, and the antigen is emulsified. Four BALB / C mice were taken, and blood was taken from the orbit before immunization as negative serum. For the first immunization, CAP-OVA was emulsified with an equal volume of complete Freund's adjuvant and injected intraperitoneally at a dose of 100 μ / mouse; 2 weeks later, CAP-OVA After mixing with Freund's incomplete adjuvant, they were immunized again; after that, boosted immunization once every other week, and then incomplete Freund's adjuvant was injected sequentially in the tail vein. Three days before the fusion, booster immunization was performed once, and the mice were slowly and directly injected with ...

Embodiment 3

[0051] Example 3 Biotinylation of Chloramphenicol-specific Monoclonal Antibody

[0052] Take 1 mg of chloramphenicol monoclonal antibody and dissolve it with 45 μL of 10 mM Sulfo-NHS-Biotin. Stir the reaction for 4-5 hours at room temperature to fully biotinylate the antibody. Then pass through YM-10 column with 0.1mol / L, PBS of pH7.4, replace the buffer solution and remove unreacted small molecule BNHS. Aliquot various biotinylated antibodies in aliquots and store in a freezer at -20°C.

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Abstract

The invention belongs to the technical field of biological engineering, and relates to a liquid-phase chip kit used for detecting chloromycetin in an aquatic product. The method comprises the steps that: (1) a chloromycetin artificial antigen is prepared; (2) chloromycetin monoclonal antibody is prepared and purified; (3) the artificial antigen obtained in the step (1) is used for coupling liquid-phase chip fluorescent microspheres; (4) the monoclonal antibody obtained in the step (2) is used for biotinylation; and (5) the coupled microspheres obtained in the step (3) and the biotinylated antibody obtained in the step (4) are used for detecting chloromycetin in an aquatic product through the liquid-phase chip method. The method provided by the invention can be used for detecting chloromycetin residue index with high speed, good specificity, and high specificity. A chloromycetin detection limit is lower than 0.1ng / L.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for detecting chloramphenicol in aquatic products by a liquid phase chip. Background technique [0002] Chloramphenicol (Chloramphenicol, CAP) is a class of broad-spectrum antibiotics extracted from Streptomyces venezensee. It is also widely used in animal husbandry at home and abroad. However, with the in-depth research on the clinical application of chloramphenicol, it is found that long-term micro-intake of chloramphenicol not only produces drug resistance in Salmonella and Escherichia coli, but also causes the imbalance of normal flora in the body and inhibits bone marrow function. , leading to related diseases such as aplastic anemia and other malignant blood diseases, seriously endangering human health. In recent years, countries such as the United States, the European Union, and Japan have all stipulated that chloramphenicol is prohibited from being used in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 陈政晓李琦明王艳李健乔元彪陈沁
Owner SHANGHAI UNIV
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