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A technology of enzyme-linked immunosorbent reagents and sulfa drugs, which is applied to the detection of sulfa drug residues in food and feed. High, long detection time and other issues
Inactive Publication Date: 2008-01-30
JIANSGU INST OF MICROBIOLOGY
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But the disadvantage is that the pretreatment steps such as sample extraction and purification are cumbersome, the cost of detection is high, and the requirements for operators are also high. It is not suitable for screening and detection of large batches of samples, and it is difficult to promote.
Microbiological methods lack sensitivity and specificity, cumbersome preparation work, long detection time, difficulties in both qualitative and quantitative aspects, and cannot meet the requirements of determination
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Embodiment 1
[0027] 1. Preparation of immune antigen, antibody and enzyme-labeled antigen
[0028] 1. Preparation of immune antigen
[0029] The artificial immune antigen of the present invention is a sulfonamide drug-carrier protein conjugate prepared by a diazo coupling method, and the sulfa drug is sulfamethazine.
[0030] Weigh 20mg ~ 100mg sulfamethazine (SMZ), add 0.5mol / L sulfuric acid, stir overnight at 4°C in the dark, place in an ice bath, add 1.9% sodium nitrite solution dropwise, and react in the dark for about 20 minutes, add 180 μL ammonium sulfamate solution, the reaction was terminated with diazotized sulfamethazine solution.
[0031] The carrier protein is bovine serum albumin (BSA). Weigh 30mg~600mg of bovine serum albumin, dissolve it in 6mL of 0.2mol / L pH10 carbonate buffer, then add the above-mentioned diazotized sulfamethazine solution, adjust the pH of the reaction solution to 9-10, and store it at 4°C. Stir lightly for 18 hours. The reaction solution was dialyze...
Embodiment 2
[0083] The antigen was changed to sulfonamide standard substance, the antibody was changed to anti-sulfonamide antibody solution, the enzyme-labeled antigen was changed to horseradish peroxidase-sulfonamide, the reaction plate was changed to anti-sulfonamide antibody coating, and the preparation of the remaining reagent bottles was the same as in Example 1, namely It is a sulfonamide enzyme-linked immunoassay kit.
Embodiment 3
[0085] The antigen was changed to sulfamethazine standard substance, the antibody was changed to anti-sulfamethazine antibody solution, the enzyme-labeled antigen was changed to horseradish peroxidase-sulfamethazine, the reaction plate was changed to anti-sulfamethazine antibody coating, and the rest of the reagent bottles The preparation is the same as in Example 1, which is the sulfamethazine ELISA kit.
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Abstract
The invention relates to a sulfa drugs direct competition enzyme-linked immunity detection reagent box, which belongs to the enzyme-linked immunity and the veterinary drug residues detection technical fields. The invention comprises the preparation of an artificial immune antigen which can produce a sulfa drugs antibody, an enzyme label antigen, a sulfa drugs antibody, a standard liquor reagent, a bottom content liquor reagent, a display color liquor reagent, a termination liquor reagent, an enzyme label antigen diluent reagent, a packet sulfa drugs antibody porous polystyrene micro reaction board and a concentration washing liquid reagent. The invention mainly adopts the direct competition enzyme-linked immunity method to detect qualitatively or quantitatively the sulfa drugs residues quantity in the food and feed; the detection limit is larger than 2 Mug/kg, the detection time only need 2 hours; the average recovery is 70 percent to 120 percent, the errors in a batch are smaller than 10 percent, the errors between the batches are smaller than 20 percent, and the invention is characterized by simpleness, rapidness and accurateness, which can be directly used for the detection of the sulfa drugs residues quantity in the food and feed.
Description
technical field [0001] The invention relates to a sulfonamide drug direct competition enzyme-linked immunoassay kit, in particular to the detection of sulfa drug residues in food and feed, and belongs to the technical field of enzyme-linked immunosorbent immunoassay and veterinary drug residue detection. Background technique [0002] Because sulfonamides have a broad antibacterial spectrum, can inhibit most Gram-positive bacteria and negative bacteria, are stable in nature, convenient for long-term storage, relatively cheap in price, and have the advantages of various preparations such as long-acting, medium-acting, and short-acting, etc., not only It is widely used in human beings and also in veterinary clinics, and has remarkable curative effects in the prevention and treatment of diseases in animals. They can be administered in the form of medicated feed, liquid, pill or injection. Sulfonamides are usually used in poultry to prevent coccidiosis, cholera and infectious rh...
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