Colloidal gold test strip, kit and detection method for detecting T-2 toxin
A colloidal gold test paper, T-2 technology, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of expensive instruments, unable to meet the needs of rapid detection, etc., and achieve low detection limit, high sensitivity, and linear range wide effect
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Embodiment 1
[0080] Preparation of step 1 colloidal gold nanoparticles
[0081] Soak the stirring paddle and glassware in the acid tank overnight and clean; assemble the stirring paddle to the stirrer, turn on the electric heating mantle to preheat for 5 minutes; after the water in the flask boils, add 1.5mL of 1% sodium citrate .
[0082] Then turn on the stirrer to make it mix evenly, adjust the rotating speed to 500rpm, and quickly add 1mL of 1% chloroauric acid. After the solution turns wine red, remove the heat source and continue stirring; stop stirring when the temperature drops to about 50°C, take the colloidal gold and use a volumetric flask to set the volume to 100mL.
[0083] Step 2 Preparation of T-2 toxin monoclonal antibody-colloidal gold label
[0084] Take 1mL of colloidal gold, add 20μL of 0.1M sodium carbonate to adjust the pH to about 8.5, add 4μg / mL T-2 toxin monoclonal antibody, mix for 30min, add 120μL of 10% BSA, block for 15min, centrifuge at 10000rpm for 15min, d...
Embodiment 2
[0086] The preparation of embodiment 2 test strips
[0087] Prepare T-line antigen streak solution: PB solution containing 1.5mg / mL T-2 toxin complete antigen (T2-BSA), 2.5wt% sucrose, 50mM, pH value is 8.5;
[0088] Prepare C-line antibody streak solution: containing 0.3mg / mL goat anti-mouse antibody, containing 2.5wt% sucrose, diluted with 50mM PB solution, and the pH value is 8.5.
[0089] The T-line antigen streak solution and the C-line antibody streak solution were respectively streaked and coated on the NC membrane. The streaking flow rate was 1 μL / cm, and the distance between the T line and the C line was 5 mm. Place the scratched NC film in an oven at 37°C for 4 hours and take it out.
[0090] Spray the colloidal gold labeled with the T-2 toxin monoclonal antibody prepared in Example 1 on the binding pad at a rate of 5 μL / cm, repeat the spraying for each binding pad 3 times, and place it in an oven at 37°C for 6 hours after completion. ; Paste sample pads, binding p...
experiment example 1
[0097] 1. Optimization of colloidal gold nanoparticles
[0098] Soak the stirring paddle and glassware in the acid tank overnight and clean; assemble the stirring paddle on the stirrer, turn on the electric heating mantle to preheat for 5 minutes; after the water in the flask boils, add 1.5mL, 1.6mL, 1.7mL , 1.8 mL, 1.9 mL of 1% sodium citrate to optimize the stability of the synthesized colloidal gold.
[0099] Then turn on the stirrer to make it mix evenly, adjust the rotating speed to 500rpm, and quickly add 1mL of 1% chloroauric acid. After the solution turns wine red, remove the heat source and continue to stir; stop stirring when the temperature drops to about 50°C, take the colloidal gold and use a volumetric flask to set the volume to 100mL, and measure the ultraviolet absorption peak. The result is as figure 1 shown.
[0100] Depend on figure 1 It can be seen that the maximum absorption peak of the colloidal gold synthesized by adding 1.5mL of 1% sodium citrate is...
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