Anti-IBDV-ELISA-Ab
For the detection of antibodies to infectious bursal disease virus (Gumboro disease)
General provisions
The kit is intended for the determination of antibodies to infectious bursal disease virus (Gumboro disease) in chicken blood serum by indirect solid-phase enzyme-linked immunosorbent assay (ELISA).The kit is designed for 96 tests.
Kit contents
The kit includes the following components:- Dismountable plate with immobilized IBDV antigens (1 pc.)
- Positive control sample (1 vial of 1 mL volume)
- Negative control sample (1 vial of 1 mL volume)
- Dilution solution (1 vial of 20 mL volume)
- Incubation solution (1 vial of 20 mL volume)
- Chromogen solution (1 vial of 12 mL volume)
- Washing buffer 20x (1 vial of 30 mL volume)
- Stop solution (1 vial of 12 mL volume)
Storage conditions and shelf life
Shelf life is 12 months from the date of manufacture when stored in a dry, dark place at a temperature of 2°C to 8°C.Brief outline of the analysis setup
| № п/п | Step | Description | Temperature, duration | Notes |
|---|---|---|---|---|
| 1 | Preparation of components | All components must be kept at room temperature | At least 60 min at a temperature of +18°С …+25°С | |
| 2 | Preparation of 1x washing solution | Preparation of 1x washing solution (see section 5.4) | +18°С …+25°С | |
| 3 | Sample preparation | Dispense 190 μL of dilution solution (DS) into the wells of the sample dilution plate, add 10 μL of the test samples, and mix thoroughly by pipetting (see section 5.3.2) | +18°С …+25°С | Adding the sample causes a change in the shade of the solution in the well |
| 4 | Addition of incubation solution (IS) | Dispense 190 μL of incubation solution (IS) into the immunosorbent wells designated for the analyzed samples | Addition time must not exceed 15 minutes | |
| 5 | Addition of positive control (PC) and negative control (NC) | Dispense 100 μL of PC and NC control samples into the immunosorbent wells | After addition, mix the contents of the wells by tapping the edge of the plate for 30 seconds! | |
| 6 | Addition of pre-prepared samples | Dispense 10 μL of the pre-prepared samples into the plate wells containing IS, and mix by pipetting | ||
| 7 | Incubation | Seal the plate wells with a film and place for incubation | 30 minutes at a temperature of +18°C...+25°C | |
| 8 | Washing | 300 μL of 1x washing solution per well (5 times) with shaking for 5 sec. between filling and emptying the wells | +18°С …+25°С | When using an automatic plate washer, carefully monitor the accuracy of filling and the completeness of liquid removal from the microplate wells |
| 9 | Addition of conjugate solution | Dispense 100 μL of conjugate (C) into all wells of the plate | +18°С …+25°С | |
| 10 | Incubation | Seal the plate wells with a film and place for incubation | 30 minutes at a temperature of +18°C...+25°C | |
| 11 | Washing | Same as step 8 | ||
| 12 | Addition of chromogen solution (CS) | Dispense 100 μL of chromogen solution (CS) into all wells of the plate | +18°С …+25°С | |
| 13 | Incubation | Place the plate for incubation | In a place protected from light | |
| 14 | Addition of stop solution (SS) | Dispense 100 μL of stop solution (SS) into all wells of the plate | 15 minutes at a temperature of +18°C...+25°C | |
| 15 | Result recording | Measure the optical density (OD): main filter — 450 nm | The time between stopping the reaction and result recording must not exceed 30 min |