Enzyme linked immunosorbent assay kit for detecting 21 sulfonamides and 11 quinolones and application of enzyme linked immunosorbent assay kit
An enzyme-linked immunosorbent reagent and drug technology, which is applied in the directions of microorganism-based methods, immunoglobulins, biochemical equipment and methods, etc., to achieve the effects of simple sample pretreatment process, strong practicability, and simple and easy testing methods.
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Embodiment 1
[0064] Embodiment 1, the preparation and application of the enzyme-linked immunosorbent assay kit that detects sulfa drug and / or quinolone drug
[0065] 1. Preparation of an enzyme-linked immunosorbent assay kit for detecting sulfonamides and / or quinolones
[0066] ELISA kits for the detection of sulfonamides and / or quinolones include:
[0067] (1) Pre-coated ELISA plate: ELISA plate coated with the original coating (the conjugate of sulfamethazine and carrier protein, and the conjugate of norfloxacin and carrier protein);
[0068] (2) Primary antibody: anti-sulfamethazine antibody working solution, and anti-norfloxacin antibody working solution, that is, anti-sulfamethazine antibody and anti-norfloxacin antibody are respectively diluted with antibody (0.02M phosphate buffer containing 0.05% Triton X-100 and 0.02% Proclin300) diluted to a protein concentration of 0.1 μg / mL.
[0069] (3) Enzyme-labeled secondary antibody: Alkaline phosphatase-labeled goat anti-mouse secondary...
Embodiment 2
[0152] Example 2, the ELISA kit precision, accuracy, specificity and shelf life experiments prepared in Example 1
[0153] In this example, the precision, accuracy, specificity, and shelf life of the enzyme-linked immunosorbent kit prepared in Example 1 will be tested. For the use method and result analysis of the kit involved, please refer to Step 2 of Example 1. step.
[0154] 1. Kit precision experiment
[0155] 1. Standard product precision test
[0156] Get three batches from the enzyme-linked immunosorbent assay kit prepared in embodiment 1 and carry out precision test, each batch respectively extracts 10 kits, each plate each extracts 20 microwells, measures 20 μ g / L standard solution (sulfamethazine standard product and ciprofloxacin standard solution, both concentrations are 20μg / L) absorbance value, calculate the coefficient of variation (CV%), the results are shown in Table 1.
[0157] Table 1 Standard precision test results (CV%)
[0158]
[0159] From the t...
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