1. Brief Introduction

The BIOKAU® IBDV kit use IBDV solid phase antigen to detect specific antibody against IBDV antigen, have high sensitivity, specificity, reproducibility, and is easy to operate quickly and can be used dividually in multiple times.

2. Reagents

1)         Antigen coated microplate................................................ 1 piece (96 wells)

2)         HRP Conjugate solution........................................... 1 bottle (11 mL/bottle)

3)         Dilution buffer.......................................................... 1 bottle (20 mL/bottle)

4)         Substrate A /B........................................................ 1 tube each (7 mL/tube)

5)         Washing buffer(26x concentrated)............................. 1 bottle (20 mL/bottle)

6)         Negative control serum................................................ 1 tube (0.7 mL/tube)

7)         Positive control serum................................................. 1 tube (0.7 mL/tube)

8)         Stop solution................................................................. 1 tube (7 mL/tube)

3. Warning and Procautions

1.         All reagents should be adjusted to the room temperature before using the test kit.

2.         Never mix up reagents from test kits in different batch number.

3.         Prevent the microplate to get damp.

4.         Keep dry of remaining microplates after use

4. Preparation

Dilute 26× concentrated washing solution at 1: 25 with distilled water.

5. Test Procedure

1.         Take out the pre-coated microplate for at least 30 min to room temperature (20-25 ℃).

2.         Put the required micro-well strips into plate frames. Re-sealed the unused microplate, store at 2-8 ℃, not frozen.

3.         Set 2 wells for Negative control serum, Positive control serum and blank control wells separately.

4.         Add 100 μL Negative/Positive control serum to its wells; Don’t add any to the 2 blank contril wells; Dilute the sample at 1:100 with the Dilution buffer and add to the wells, 100 μL each.

5.         Shake softly, incubate at 37℃ for 30 min.

6.         Pour the liquid out of the wells, add 300 μL diluted washing solution to each well, static for 1 min, pour out. Repeat 5 times, then pat to dry on absorbent paper.

7.         Add 100 μL HRP Conjugate solution(except blank wells) to each well, and incubate at 37℃ for 30 min.

8.         Repeat the step 6(washing).

9.         Add substrate A and B one drop(50 μL) respectively to each well, mix properly, react for 10 min at room temperature at dark.

10.     Add stop solution one drop (50 μL) in each well, and measure the result within 10 min.

6. Results

Set zero for the blank well, and test the OD450nm (recommend 450nm and 630 nm )value on the microplate-reader. The conditions for the test to be tenable are that the positive control wells’ average OD450nm value is greater than or equal to 0.4, and the negative control wells’ average OD450nm value is less than 0.1.(all value should minus the average value of blank control)

Cut-off value = 0.15 + A(absorbance of negative control)

Interpretation of results:

A450 (patient) ≥ cut-off: positive

A450 (patient) ＜ cut-off: negative

Specifications: 96 wells/kit.

Expiry date: 12 months.

Storage: Storing at 2-8℃, in the dark.