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Fluorescence immunoassay chromatography test paper for cefalexin residue and preparation of test paper

A technology of fluorescence immunochromatography and cephalexin, which is applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of rapid detection of unsuitable samples, high cost, cumbersome measurement operation, etc., and achieve simple measurement method, simple processing process and high sensitivity high effect

Active Publication Date: 2014-07-02
CHINESE ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

Although chromatography or chromatography-mass spectrometry has high sensitivity, the sample pretreatment and determination operations are cumbersome and expensive, and are not suitable for rapid detection of a large number of samples

Method used

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  • Fluorescence immunoassay chromatography test paper for cefalexin residue and preparation of test paper

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Embodiment 1

[0034] Embodiment 1: Preparation of cephalexin residual fluorescent immunochromatographic detection test strip

[0035] 1. Synthesis of Protein-Coupled Antigens

[0036](1) Synthesis of cephalexin immune antigen

[0037] The immune antigen was obtained by coupling cephalexin and bovine serum albumin (BSA) by carbodiimide method.

[0038] The specific operation is as follows: Dissolve 15 mg of cephalexin in 5 mL of PBS (0.01 mol / L, pH 7.4), add 25 mg of carbodiimide (EDC) and 10 mg of N-hydroxysuccinimide (NHS) and stir for 30 minutes, and take 10 mg of BSA Dissolve in activated cephalexin solution, react with stirring at 25°C for 2 hours, then react overnight at 4°C. Dialyzed with PBS for 3 days at 4°C to obtain the cephalexin immune antigen, which was subpackaged and frozen.

[0039] (2) Synthesis of cephalexin-coated antigen

[0040] Coupling of cephalexin and ovalbumin (OVA) was carried out by glutaraldehyde method to obtain the coated antigen.

[0041] Concrete operat...

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Abstract

The invention discloses a piece of fluorescence immunoassay chromatography test paper which is rapid, sensitive, simple and convenient to operate to test the residual quantity of cefalexin, and preparation of the test paper. The test paper comprises a sample pad, a binding pad, a nitrocellulose membrane and a water absorbing pad, which are adhered to a substrate in a mutual lap joint manner in sequence, wherein the nitrocellulose membrane is coated with a detection line and a quality control line; the binding pad is coated with a cefalexin monoclonal antibody with a fluorescent mark. The preparation method of the fluorescence immunoassay chromatography test paper for cefalexin residue comprises the following steps: synthesizing cefalexin-ovalbumin coating antigen, preparing a goat anti-mouse immune globulin antibody, coating the cefalexin-ovalbumin coating antigen and the goat anti-mouse immune globulin antibody onthe nitrocellulose membrane to serve as the detection line and the quality control line, preparing a fluorescence nano-particle mark cefalexin monoclonal antibody, then coating glass fiber to serve as the binding pad, sequentially adhering the sample pad, the binding pad, the nitrocellulose membrane and the water absorbing pad to a back plate in the lap joint manner in sequence, cutting the obtained test paper into the width of 4mm, and preserving at normal temperature.

Description

technical field [0001] The invention relates to a rapid fluorescence immunoassay technology for veterinary drug residues, in particular to a fluorescence immunochromatographic detection test strip for detection of cephalexin residues and its preparation. Background technique [0002] Cephalexin is a β-lactam antibiotic. It is widely used in veterinary clinics 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 residue in animal products and cause serious harm to human health. Regarding the residues of the drug in food, the Ministry of Agriculture issued the "Maximum Residue Limits of Veterinary Drugs in Animal Foods", which stipulates that the maximum residue limit in milk is 100 μg / kg, and the maximum residue limit in muscle and fat is 200 μg / kg . [0003] At present, the methods used to d...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/54373G01N33/558G01N33/582
Inventor 邹明强张帆陈艳胡佳丽
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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