Description
This HCV ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of HCV in the sample, this HCV ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a cutoff value. The existence or not of HCV in the samples is then determined by comparing the O.D. of the samples to the CUT OFF.
ASSAY PROCEDURE
- Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
- Separately add Positive control and Negative control 50μl to the Positive and Negative well; Add testing sample 10μl then add Sample Diluent 40μl to testing sample well.
- Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
- Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400 μ l) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.
- Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.
- Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate toensure thorough mixing.
- Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.
CALCULATION OF RESULTS
- Test validity: the average of Positive control well≥1.00; the average of Negative control well ≤0.15.
- Calculate Critical (CUT OFF): Critical= the average of Negative control well + 0.15.
Negative Result: sample OD< Calculate Critical (CUT OFF) is Negative.
Positive Result: sample OD≥ Calculate Critical (CUT OFF) is Positive.
Specification:96T
