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Three-in-one colloidal gold chromatography test strip for detecting thiamphenicol, chloramphenicol and florfenicol and its preparation method

A technology of thiamphenicol and florfenicol, which is applied in the field of immunological detection, can solve the problems of high cost, high detection cost, unsuitable for low-cost detection of enterprises, etc., and achieve the effect of fast detection and easy operation

Active Publication Date: 2017-05-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Instrumental analysis methods mainly include high performance liquid chromatography, liquid chromatography-mass spectrometry (HPLC-MS), gas chromatography-mass spectrometry (GC-MS), etc. These methods have high precision but require expensive instruments and equipment, professional and technical personnel are required for operation, and Cannot adapt to high-throughput, on-site rapid detection
Immunoassays are mainly enzyme-linked immunoassays, which have high testing costs and are not suitable for low-cost testing needs of enterprises.

Method used

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  • Three-in-one colloidal gold chromatography test strip for detecting thiamphenicol, chloramphenicol and florfenicol and its preparation method
  • Three-in-one colloidal gold chromatography test strip for detecting thiamphenicol, chloramphenicol and florfenicol and its preparation method
  • Three-in-one colloidal gold chromatography test strip for detecting thiamphenicol, chloramphenicol and florfenicol and its preparation method

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Experimental program
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Effect test

Embodiment 1

[0025] Such as figure 1 As shown, a three-in-one colloidal gold immunochromatographic test strip for detecting thiamphenicol, chloramphenicol and florfenicol, including polyvinyl chloride PVC backing 1, and the front end of polyvinyl chloride backing 1 is set The sample pad 2, the sample pad 2 is connected to the front end of the nitrocellulose membrane 4 through the binding pad 3, and the rear end of the nitrocellulose membrane 4 is connected to the water-absorbing pad 5; the binding pad 3 contains a colloidal gold-labeled monoclonal antibody; the nitrocellulose The prime film 4 is coated with chloramphenicol sodium succinate-BSA detection line T line 6 and goat anti-mouse IgG control line C line 7 .

Embodiment 2

[0027] The preparation method of the three-in-one colloidal gold immunochromatography test strip for detecting thiamphenicol, chloramphenicol and florfenicol, the steps are:

[0028] (1) Preparation and purification of monoclonal antibodies that can simultaneously recognize thiamphenicol, chloramphenicol and florfenicol: the analogue of thiamphenicol, namely 1-(4-aminophenyl)-2- Dichloroethylamine-1,3-propanediol (TAP-NH 2 ) as a hapten, coupled with bovine serum albumin (BSA) by the diazotization method to obtain an immunogen, mixed with Freund's adjuvant to immunize BALB / c mice; take the spleen cells and myeloma cells of qualified immunized mice Sp2 / 0 fusion, after three times of subcloning and indirect competition ELISA screening, a cell line that can stably secrete monoclonal antibodies that can recognize thiamphenicol, chloramphenicol and florfenicol at the same time was obtained. After expanding the culture, Inject into mice to induce ascites; use octanoic acid-saturate...

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Abstract

The invention provides a three-in-one colloidal gold chromatographic test strip for detecting thiamphenicol, chloramphenicol and florfenicol and a preparation method thereof, and belongs to the technical field of immunological detection. The preparation method comprises two parts, namely, the preparation of a monoclonal antibody and the preparation of a colloidal gold chromatographic test strip, wherein the monoclonal antibody can be used for recognizing thiamphenicol, chloramphenicol and florfenicol at the same time and is high in sensitivity; the colloidal gold chromatographic test strip comprises a polyvinyl chloride backing; a sample pad is arranged at the front end of the polyvinyl chloride backing and is connected with the front end of a nitrocellulose membrane; the rear end of the nitrocellulose membrane is connected with a water absorbing pad; the monoclonal antibody marked by colloidal gold is used as a combining pad; the nitrocellulose membrane is sequentially wrapped with a chloramphenicol sodium succinate-BSA (Bovine Serum Albumin) antigen test ray T and a goat-anti-mouse IgG control ray C. The three-in-one colloidal gold chromatographic test strip for detecting thiamphenicol, chloramphenicol and florfenicol is fast to detect, and the detection needs only 3 to 5 minutes; the test strip is convenient to carry, and suitable for detection on site; the operation is simple and convenient and does not need a professional technical person.

Description

technical field [0001] The invention relates to a three-in-one colloidal gold immunochromatographic test strip for detecting thiamphenicol, chloramphenicol and florfenicol and a preparation method thereof, belonging to the technical field of immunological detection. Background technique [0002] Chloramphenicol antibiotics are one of the widely used veterinary drugs. At present, there are mainly three drugs: chloramphenicol (CAP), thiamphenicol (TAP) and florfenicol (FF). Among them, thiamphenicol is a derivative of florfenicol, and their antibacterial effect is similar to that of chloramphenicol. Elements are similar. Chloramphenicol can cause aplastic anemia in humans. This toxic effect has nothing to do with the dose and course of treatment. The use of chloramphenicol has been banned or strictly restricted internationally. Florfenicol and thiamphenicol are substitutes for chloramphenicol, but their extensive use leads to a large amount of residues in animal products and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 匡华郭玲玲胥传来徐丽广马伟刘丽强宋珊珊吴晓玲
Owner JIANGNAN UNIV
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