Biosensor for detecting oxytetracycline
A biosensor, oxytetracycline technology, applied in the field of biosensors, can solve the problems of long detection period, high cost, low specificity and sensitivity, etc., and achieve the effects of short detection period, simple operation and good sensitivity
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Embodiment 1
[0030] Example 1 Preparation of gold nanometers.
[0031] (1) Add 200mL ultrapure water into the three-neck flask;
[0032] (2) Take 500uL of HAuCl with a concentration of 0.04g / mL 4 In the centrifuge tube, add 200mL ultrapure water, stir and heat to boil, stirring speed 450rpm;
[0033] (3) Under stirring conditions, take 3ml of 1% trisodium citrate solution and quickly add it to the solution in step (2). The color of the solution changes from light yellow to wine red. After continuing to heat for 15 minutes, remove the heat source and Slowly cool to room temperature, and store at 4°C for later use.
[0034] According to the absorbance value at 530 nm, the concentration of gold nanoparticles in the above solution is about 0.3 nM.
Embodiment 2
[0035] Example 2 Modification of gold nanometers.
[0036] (1) Take 1 mL of the nano-gold solution in Example 1 in a centrifuge tube, centrifuge for 10 min until the supernatant is colorless and transparent, remove the supernatant, add 300 μL of sterilized water to concentrate the nano-gold solution to 3 nM;
[0037] (2) Transfer the above solution into a 1 mL glass bottle, seal it with tinfoil, and place it at room temperature for 30 min, then add 150 μL of S1 with a concentration of 30 μM, mix well, and place it at 4 °C for 24 h;
[0038] (3) To the above solution, slowly add 50 μL of PB buffer several times, add magnets (soaked in aqua regia the day before and rinse repeatedly with sterile water), stir for 10 min, then continue to add 27 μL of PBS buffer, take out the magnets Place it at 4°C for 48 h;
[0039] (4) Transfer the above solution to a centrifuge tube, add sterilized water to 1 mL, centrifuge for 10 min, remove the supernatant; then add 1 mL of sterilized water ...
Embodiment 3
[0040] Example 3 The effect of different concentrations of complete antigen on the detection of oxytetracycline.
[0041] (1) 2 μL of different concentrations of oxytetracycline complete antigen (OTC-BSA, the final concentration is 0.0001 mg / mL, 0.001 mg / mL, 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 10 mg / mL ) and 2 μL oxytetracycline solution (1 x 10 -3 mg / L) into a centrifuge tube, shake for 30 seconds, and place in a water bath at 37 degrees Celsius for 0.5 hours.
[0042] (2) Take out the mixed solution from the water bath, add 20 μL of S1-modified nano-gold solution, and put it in a water bath at 37°C for 30 minutes.
[0043] (3) After 30 minutes, take out the mixed solution from the water bath; measure the absorbance value of the mixed solution at 520 nm with a UV-visible spectrophotometer.
[0044] Take the OTC-BSA concentration as the abscissa, and take the absorbance value as the ordinate, as figure 2 , it can be seen from the figure that the detected peak UV absorption in...
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