Carbon quantum dot fluorescence immunoassay rapid detection kit for six types of veterinary drug antibiotics in poultry tissues and detection method thereof
A technology of detection kits and carbon quantum dots is applied in the field of immunodetection to achieve the effects of saving detection time, reducing costs and contracting manpower
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Embodiment l
[0029] Embodiment 1 (preparation embodiment)
[0030] Preparation of carbon quantum dots
[0031] (1) Synthesis of carbon quantum dots: 3 g of citric acid and 5 g of urea were added to 10 mL of ultrapure water to form a transparent solution. The solution was then heated in an 800W microwave oven for 5 minutes, during which time the solution turned from a colorless liquid to brown and finally to a dark brown clustered solid. After cooling to room temperature, 20 mL of water was added to dissolve the product.
[0032] (2) Purification of carbon quantum dots: After the dissolved product is centrifuged at 5000rpm for 10min, it is packed into a 3500Da dialysis bag, and dialyzed with water at room temperature overnight to obtain a brown fluorescent solution. The fluorescence spectrum is as follows: figure 1As shown, under the excitation of 365nm excitation light, there is fluorescence at 400nm-600nm.
Embodiment 2
[0033] Embodiment 2 (preparation embodiment)
[0034] (1) Coating antigen: Dilute furaltadone, nitrofurazone, nitrofurantoin, furazolidone, chloramphenicol and florfenicol coating antigen to 0.25, 0.25, 0.25, 0.25, 0.5 and 0.5 μg with pH 9.6 sodium carbonate buffer / mL, take 100-300 μL into the corresponding cuvette, and incubate overnight at 4°C.
[0035] (2) Wash the cuvette: discard the reaction solution the next day, add 200-500 μL phosphate washing solution, shake for 2 minutes, shake off the phosphate washing solution, repeat this process 3 times, and finally pat the cuvette dry on filter paper.
[0036] (3) Blocking: add 200 μL of casein blocking solution to the cuvette, incubate at 37° C. for 1.5 h, discard the casein blocking solution, and store at 4° C. after being vacuum-sealed.
Embodiment 3
[0037] Embodiment 3 (application embodiment)
[0038] Application technology of carbon quantum dot fluorescent immunoassay rapid detection kit
[0039] (1) Add standard solution and horseradish peroxidase-labeled antibody working solution: Add 100 μL of the standard dilution solution containing a certain concentration of the analyte to the cuvette coated with the corresponding analyte-coated antigen, and then add 100 μL of the analyte Add the enzyme-labeled antibody mixture corresponding to the test object into each reaction cup, two parallels for each concentration, incubate at 37°C for 20 minutes, discard the reaction solution, add 200-500 μL phosphate washing solution, shake for 2 minutes, and shake off the phosphate washing solution , and so repeated 3 times, and finally the plate was patted dry on filter paper.
[0040] (2) Color development: 50 μL each of carbon quantum dot fluorescent reaction solution A and carbon quantum dot fluorescent reaction solution B were added...
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