Test strip for detecting tetanus antibody based on quantum dot fluorescent microsphere chromatography and preparation method and use method thereof
A technology of fluorescent microspheres and quantum dots, which is applied in biological testing, measuring devices, and analytical materials to achieve fast detection speed, good stability, and reduced material interference
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0031] The preparation of quantum dot fluorescent microsphere-labeled protein, the steps are as follows:
[0032] (1) First, 100 μL of carboxylated quantum dot microspheres (QDNB) were centrifuged at 15,000 for 30 min, and then reconstituted in 100 μL of 50 mM pH 6.0 MES buffer. Then 15 μL of 8 mM N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) was added, and the reaction was rotated in a dark room for 30 min to activate the quantum dot microspheres. Centrifuge at 15,000 for 30 min.
[0033] (2) Use 100 μL of 50 mM pH 7.4 boric acid buffer for ultrasonic reconstitution, then add different volumes of 5 mg / ml Fc, respectively 2 μL, 3 μL, 5 μL, 7 μL, 10 μL, and rotate in a dark room for 60 min. Centrifuge at 15000, 30min.
[0034](3) Add 100 μL of 2% BSA blocking solution to reconstitute and perform blocking treatment. After 50 min of reaction in the dark room, centrifuge at 15,000 for 30 min. The final prepared QDNB-Fc is stored in 100 μL of boric acid buffer...
Embodiment 2
[0037] The preparation method of the test strip for the detection of tetanus antibodies based on quantum dot fluorescence microsphere chromatography, the steps are as follows: first seal the sample pad with a mass concentration of 0.5-5% TW20 solution, and then draw the detection line and In the quality control line, the treated sample pad and NC film were vacuum-dried at 37°C for 1 hour, and then the sample pad, NC film and water-absorbing pad were pasted on the PVC base plate in turn, overlapped by 2-3mm, and finally cut into 3.9mm width, the finished test strip is obtained.
[0038] Detection of tetanus antibodies by fluorescent microsphere immunochromatographic test strips:
[0039] Prepare the test strip prepared by the above steps, prepare a tetanus antibody standard solution with a certain concentration gradient (0-0.022IU), mix the concentration gradient tetanus antibody standard solution with a certain amount of QDNB-Fc, and take the solution. 90 μL of the mixed solu...
Embodiment 3
[0041] Interference of different types of antibodies on tetanus antibody detection:
[0042] The complex of the quantum dot microsphere antibody prepared by the above steps and the test strip coated with the antigen are then prepared with a tetanus antibody standard solution concentration of 0.01 IU, and the concentration of the interfering antibody is 10 equivalents. For example, the concentration of pertussis antibody is 0.1 IU, 0.1 IU for rabies antibody, 5 μg / ml for cytokeratin antibody, 5 μg / ml for thyroid transcription factor antibody, mixed solution of 0.1 IU pertussis antibody and 0.01 IU tetanus antibody, respectively. After the sample was mixed with a certain amount of QDNB-Fc, 90 μL of the mixed solution was dropped on the sample pad of the tetanus antibody detection test strip. After 10 minutes, the fluorescence peak intensity corresponding to different antibodies was recorded, and each group of experiments was repeated 3 times in parallel. like Figure 4 As shown...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More