Method for quickly immunologically detecting beta-lactamase in milk

A lactamase and rapid immunization technology, applied in the field of ELISA detection of beta-lactamase, can solve the problem of lack of immune detection of beta-lactamase, etc., and achieve the effect of ensuring food safety, high accuracy and low cost

Inactive Publication Date: 2010-05-19
BIOLOGY INST OF HEBEI ACAD OF SCI
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Problems solved by technology

There is currently no method for the accurate, specific and rapid immunodetection of β-lactamase illegally added to milk

Method used

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  • Method for quickly immunologically detecting beta-lactamase in milk
  • Method for quickly immunologically detecting beta-lactamase in milk

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Embodiment Construction

[0017] In order to implement the present invention better, the preferred embodiments are as follows.

[0018] BALB / c mice and New Zealand white rabbits were immunized with β-lactamase respectively, and mouse anti-β-lactamase monoclonal antibody and rabbit anti-β-lactamase polyclonal antibody were prepared. The titer of the mouse anti-β-lactamase monoclonal antibody is 1:13.1 million, the subtype is IgG1, and the affinity is 1.046×10 10 L / mol, negative reaction with other related substances, only positive reaction with antigen β-lactamase; rabbit anti-β-lactamase titer is 1:6.55 million, only positive reaction with antigen β-lactamase. Both monoclonal and polyclonal antibodies are purified by protein A and used. Mouse anti-β-lactamase monoclonal antibody used for coating, rabbit anti-β-lactamase polyclonal antibody, horseradish peroxidase-labeled goat anti-rabbit antibody can realize the detection of β-lactamase in raw milk Three-antibody sandwich assay. The detection sensit...

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Abstract

The invention provides a method for quickly immunologically detecting a beta-lactamase in milk, and belongs to the technical field of immunological detection. The method is characterized in that: a three-antibody sandwich method reagent consists of a rat anti-beta-lactamase monoclonal antibody coated on an elisa plate, a rabbit anti-beta-lactamase polyclonal antibody and a horseradish peroxidase labelled sheep and rabbit antibody; and immunological detection is performed to detect the beta-lactamase in a raw milk by a three-antibody sandwich method. Compared with the prior art, the method canaccurately, specially and quickly immunologically detect the beta-lactamase illegally added into the milk so as to timely monitor and control the quality of the raw milk from the source and guaranteethe safety of the milk and the health of human, and is good in social and economic efficiency.

Description

technical field [0001] The invention belongs to the technical field of immunoassay, in particular to an ELISA detection method for β-lactamase added to raw milk in the field of animal food safety or animal husbandry production. Background technique [0002] Currently, penicillin, as a β-lactam drug, is the drug of choice for the treatment of bovine mastitis and is the most common residual antibiotic in milk. Since most dairy companies in China refuse to purchase milk with excessive antibiotic residues, driven by economic interests, some illegal dairy stations artificially use some biological agents to degrade the antibiotic residues in milk in order to seek their own economic interests. The so-called artificial "antibiotic-free milk". Since 2005, several companies have publicly announced the sale of anti-antibiotics that decompose residual antibiotics in milk. The main component of the antibiotic is β-lactamase, which is produced and secreted by Gram-positive bacteria and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 闫静辉张小兵吴萌李春生邸禄芹齐颖颖李君华高志肖陈英珠程华董超李亚璞谢莉时玮
Owner BIOLOGY INST OF HEBEI ACAD OF SCI
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