Instructions for using the fluorescent quantitative PCR kit for cysticercus probe method:

The sensitivity of the assay comes from the enzyme used as the report. Enzymes are organic catalysts, and a small amount of enzyme can induce a large number of catalytic reactions, resulting in observable color reactions. Therefore, this system is often referred to as an enzyme amplification system.

1. Dilution of standard samples: This kit provides one original standard sample, and users can dilute it in a small test tube according to the following chart. Add 150 μ l of standard dilution to 150 μ l of the original standard of standard 5 at a concentration of 24 μ g/ml

Add 150 μ l of standard dilution solution to 150 μ l of standard 5, with a concentration of 12 μ g/ml

Add 150 μ l of standard dilution to 150 μ l of standard dilution solution for standard 4, with a concentration of 6 μ g/ml

Add 150 μ l of standard dilution solution to 150 μ l of standard 2 at a concentration of 3 μ g/ml

Add 150 μ l of standard dilution to 150 μ l of standard dilution solution for standard 1 at a concentration of 1.5 μ g/ml

2. Sample addition: Set up blank holes, standard holes, and sample holes for testing. Accurately add 50 μ l of the standard sample on the enzyme-linked immunosorbent assay (ELISA) coated plate, first add 40 μ l of sample diluent to the well of the test sample, and then add 10 μ l of the test sample (with a final dilution of 5 times). Add the sample to the bottom of the enzyme-linked immunosorbent assay plate well, avoiding touching the well wall as much as possible, and gently shake and mix well. |

3. Incubation: Cover the plate with a sealing film and incubate at 37 ℃ for 30 minutes.

4. Solution preparation: Dilute 30 times the concentrated washing solution with distilled water and set aside for later use

5. Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each hole with washing solution, let it stand for 30 seconds, then discard. Repeat this process 5 times and pat dry.

6. Enzyme addition: Add 50 μ l of enzyme labeled reagent to each well, except for blank wells.

7. Incubation: Follow the same procedure as in 3.

8. Washing: Follow the same procedure as in 5.

9. Color development: Add 50 μ l of color developing agent A50 to each well, then add 50 μ l of color developing agent B50, gently shake and mix well, and develop color at 37 ℃ in the dark for 15 minutes

10. Termination: Add 50 μ l of termination solution to each well to terminate the reaction (at this point, blue will turn yellow).

11. Measurement: Measure the absorbance (OD value) of each well in sequence using blank air conditioner zero and 450nm wavelength. The measurement should be conducted within 15 minutes after adding the termination solution.