Adprin immune colloidal gold detection card and preparation method thereof
A technology of colloidal gold and test cards, which is applied to measuring devices, instruments, scientific instruments, etc., to achieve accurate results and simple methods
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Embodiment 1
[0020] figure 1 , figure 2 Shown embodiment: among the figure 9 is a PVC rubber sheet; 7 is a sample pad; 8 is a colloidal gold binding pad, and the colloidal gold binding pad is coated with a monoclonal antibody colloidal gold marker; 10 is a coating film, i.e. nitrocellulose Plain membrane, the nitrocellulose membrane coated with adfuroline-BSA and goat anti-mouse IgG; 11 is a water-absorbent pad, made of water-absorbent materials such as filter paper.
[0021] On one end (sample end) of the PVC rubber plate 9, the sample pad 7 and the bonding pad 8 are adhered, and the sample pad 7 and the bonding pad 8 are in a side-by-side structure.
[0022] A nitrocellulose membrane 10 is adhered in the middle of the PVC rubber sheet 9 . On the nitrocellulose membrane 10, a goat anti-mouse IgG quality control line 3 and an adfuroline-BSA detection line 2 are set.
[0023] At the other end of the PVC rubber sheet 9, a water-absorbing pad 11 is adhered. One end of the nitrocellulose ...
Embodiment 2
[0025] The preparation of the adfuroline colloidal gold detection card is realized by the following steps:
[0026] Preparation of colloidal gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 (0) 5 mL, prepared into 500 mL 0.01% chloroauric acid aqueous solution, heated and boiled, then added 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Solution 5-7 mL, continue to stir and heat, when the color of the solution turns completely transparent purple red, stop heating after maintaining for 5 min, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;
[0027] Antibody pretreatment: Centrifuge the antibody to be labeled with adefoline at 1000 r / min, 4°C for 20 min, take the supernatant, and dilute it to 1 mg / mL with 0.01 mol / L PBS; or use 0.01 mol / L Dilute L PBS to 1 mg / ml, pass through a 0.22 µm filter;
[0028] Preparation of colloidal gold markers: Take 40 mL of...
Embodiment 3
[0034] The minimum detection limit test for adefurin
[0035] Accurately weigh 0.500 g of adfuroline antigen, dissolve it in ultrapure water and dilute to 100 mL, mark it as A 1 solution, take A 1 Add 1mL of the solution to a volumetric flask and add ultrapure water to 1000mL to form A 2 solution, take A 2 Add 1 mL of the solution to a volumetric flask and add ultrapure water to 1000 mL to form A 3 solution, the concentration of A3 solution is 5 ng / mL (5ppb). take A 3 Put 1 mL of the solution in 4 test tubes, and label them 1~4 respectively, add 9 mL, 4 mL, 1 mL, 0 mL of ultrapure water to the test tubes respectively, and make the concentration of Adfurfural antigen solution: 0.5 ng / mL, 1 ng / mL, 2.5 ng / mL, 5 ng / mL; take another empty test tube, add 5 mL of ultrapure water, and mark it as No. 5 test tube. Take the liquid in the test tubes No. 1-5 above as a sample and add it dropwise to the adofurin test card, and observe the results 5 minutes after adding the sample. The...
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