Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof
A technology of colloidal gold test paper and kanamycin, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of unfavorable promotion and use, complicated operation process, and cumbersome measurement methods, and achieve short detection time, low temperature requirements, The effect of easy operation
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[0030] Example 1: The structure of the immune colloidal gold test strip of the present invention
[0031] Such as figure 1 with figure 2 As shown, an immune colloidal gold test strip for detecting kanamycin residues, including sample pad 2, binding pad 3, nitrocellulose membrane 4, absorbent pad 5 and PVC backing 1, in the PVC backing 1. The sample pad 2, the binding pad 3, the nitrocellulose membrane 4 and the absorbent pad 5 are successively adhered to the upper part. There is an overlap between the binding pad 3 and the nitrocellulose membrane 4, and between the nitrocellulose membrane 4 and the absorbent pad 5 Overlap; the binding pad 3 is coated with anti-kanamycin monoclonal antibody-colloidal gold marker, and the nitrocellulose membrane 4 is respectively coated with kanamycin-carrier protein (bovine serum albumin) coupling The detection line T formed by the substance and the quality control line C formed by the rabbit anti-mouse IgG. Finally cut into 3mm wide strips and v...
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[0034] Example 2 (Preparation Example)
[0035] Preparation method of immune colloidal gold test strip for detecting kanamycin
[0036] 1. Preparation of kanamycin-carrier protein conjugate
[0037] Synthesis of immunogen: weigh 45 mg kanamycin sulfate and 10 mg hemocyanin (KLH), dissolve in 1.5 mL PBS, and stir on a magnetic stirrer. Dissolve 300 mg of EDC·HCl in 1 mL of PBS, adjust the pH to 7.4, and add it dropwise to the above mixed solution, stir at room temperature for 1 hour, and then stand at 4°C for 3 days. It was dialyzed against PBS for 3 days, the dialysate was changed 2 to 3 times a day, and further purified by a Sephadex G-25 column, packed in aliquots, and stored at -20°C.
[0038] Synthesis of the coating agent: Weigh 45 mg of kanamycin sulfate and 10 mg of BSA in 1.5 mL of PBS, and stir on a magnetic stirrer. Dissolve 300 mg of EDC·HCl in 1 mL of PBS, then add it dropwise to the above mixed solution, and stir and react at room temperature for 3 hours. It was dialy...
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[0057] Example 3 (application example)
[0058] Evaluation of the application of immunogold test strips for detecting kanamycin
[0059] 1. Specificity test
[0060] Prepare kanamycin, gentamicin, neomycin, streptomycin, chloramphenicol, cephalexin, sulfadimethoxine, and enrofloxacin into a sample with a concentration of 10ng / mL, and drop it on Test on the sample pad or sample hole. The test results (see Table 1) show that the kanamycin sample detection line has no color, and a red band appears on the quality control line, while gentamicin, neomycin, streptomycin, chloramphenicol, and cephalexin , Sulfadimethoxine, enrofloxacin sample test strip detection line and quality control line have red bands, which indicates that gentamicin, neomycin, streptomycin, chloramphenicol, cephalexin, sulfa Dimethoxypyrimidine and enrofloxacin have no cross-reaction with the test strip of the present invention, which shows that the test strip of the present invention has good specificity.
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