Nano-sensor for detecting antibiotics and preparation method and application of nano-sensor
A nano-sensor and antibiotic technology, applied in the fields of biochemical analysis and food safety testing, can solve the problems of inability to specifically indicate the type of antibiotics, complicated operation, etc., and achieve the effect of facilitating qualitative and quantitative judgment, solving quantitative difficulties, and shortening test time.
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Embodiment 1
[0024] Preparation of nanosensors for chloramphenicol and kanamycin detection and applications of the nanosensors:
[0025] (1) Preparation of gold nanoparticles with a diameter of 13nm: weigh 0.04726g HAuCl 4 Dissolve in a round-bottomed flask with 100mL of ultrapure water, put a magnetic stirrer, then connect a condenser tube and a stopper to the two ports of the two-necked flask, turn on the condensate, turn on the magnetic stirrer and start heating. In addition, 1.3693 g of trisodium citrate was dissolved in ultrapure water, and configured into a 120 mL solution to obtain a 38.8 mM trisodium citrate solution. When the reaction solution boils and the condensed water starts to flow back at a rate of 1 drop per second, remove the stopper, quickly add 12mL of 38.8mM trisodium citrate, and reinstall the stopper. At this time, the color of the solution will gradually change from light yellow to dark red, and then continue to heat and reflux for 15 minutes after turning dark red...
Embodiment 2
[0039] Preparation of nanosensor for neomycin detection and application of the nanosensor:
[0040] (1) Preparation of gold nanoparticles with a diameter of 5 nm: 70 mL of distilled water and 10 mL of chloroauric acid with a mass fraction of 0.1% were added to a 250 mL three-necked flask, and heated to boiling under vigorous stirring. Then, quickly add 4 mL of freshly prepared mixed solution with a mass fraction of 1% sodium citrate and 5 mL of a mixed solution with a mass fraction of 1% tannic acid to the above-mentioned boiling solution, and after reacting for 10 minutes, continue stirring to cool the solution to room temperature, Store at ℃.
[0041] (2) Modification of sulfhydryl ssDNA onto the surface of AuNPs: the present invention selects nucleic acid aptamers with high affinity and high specificity for neomycin and labeled with fluorescent molecules as recognition molecules, and their sequences are respectively:
[0042] Aptamer E specifically recognizing neomycin: 5′...
Embodiment 3
[0047] Preparation of nanosensors for detection of ampicillin and streptomycin and application of the nanosensors:
[0048] (1) Preparation of gold nanoparticles with a diameter of 30 nm: 100 mL of chloroauric acid with a mass fraction of 0.01% was added to a 250 mL three-necked flask, and heated to boiling under vigorous stirring. Then, quickly add 1 mL of freshly prepared 1% sodium citrate solution into the boiling solution, react for 15 minutes, continue stirring to cool the solution to room temperature, and store at 4°C.
[0049] (2) Modification of sulfhydryl ssDNA onto the surface of AuNPs: the present invention selects nucleic acid aptamers with high affinity and high specificity for streptomycin and ampicillin and labeled with fluorescent molecules as recognition molecules, and their sequences are respectively for:
[0050] Aptamer C that specifically recognizes streptomycin: 5′-SH-TAGGGAATTCGTCGACGGATCC-GGGGTCTGGTGTTCTGCTTTGTTCTGTCGGGTCGT-CTGCAGGTCGACGCATGCGCCG-Rox-3...
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