AffiVET® Veterinary FMD NSP Antibody ELISA Kit
Product Name: FMDV NSP 3ABC Ab Test, FMD NSP ELISA
Product Number: AFG-VE-10
Product Unit: 2 plates, 192 tests
For veterinary use only
FMDV (Foot-and-Mouth Disease Virus)
Causes vesicles in the mouth and feet of cloven-hoofed animals like bovids, suids, ovids, and caprids. Highly infectious, it was a major threat to the animal farming industry before vaccine development.
Description of Test
Detect FMDV-specific NSP 3ABC protein antibody in bovine, swine, caprine, and ovine species.
Test Method
- 96-well microtiter plate pre-coated with 3ABC protein.
- Samples added to wells form antigen-antibody complexes if 3ABC antibody is present.
- Non-specific antibodies discarded through washing.
- Blocking HRP conjugated 3ABC monoclonal antibody added, followed by washing.
- Substrate added, developing color if 3ABC antibody is present.
- Color intensity measured at OD450nm, inversely proportional to antibody quantity.
Precautions
- Storage: Store kit at 2-8°C; bring to room temperature before use.
- Handling: Stop solution is acidic; avoid contact with skin. Treat field samples as potentially infectious according to lab regulations. Dispose of materials according to local regulations.
Limitations of Test
Designed for veterinary use. Validate with different methodologies in your own lab for performance confirmation. Contact the manufacturer for non-standard use.
Reagent Provided
Kit Contents for 192 Tests
- Microtiter Plate (2 x 96 wells)
- Positive Control (1 x 1 ml)
- Negative Control (1 x 1 ml)
- Enzyme Conjugate (1 x 25 ml)
- TMB Substrate (1 x 25 ml)
- Stop Solution (1 x 20 ml, not provided due to shipping restrictions, 2M sulfuric acid to be prepared by the client)
- 25X Wash Buffer (1 x 30 ml)
- Sample Buffer (1 x 30 ml)
- Kit Instructions (1 set)
Kit Contents for 480 Tests
- Microtiter Plate: (5 x 96 wells)
Positive Control: (2 x 1 ml)
Negative Control: (2 x 1 ml)
Enzyme Conjugate: (3 x 25 ml)
TMB Substrate: (3 x 25 ml)
Stop Solution: (2 x 20 ml, not provided due to shipping restrictions, 2M sulfuric acid to be prepared by the client)
25X Wash Buffer: (3 x 30 ml)
Sample Buffer: (2 x 30 ml)
Kit Instructions: 1 set
Instruments Required
- ELISA Reader
- Micropipettes (20-200 µl and Multi-Channel 50-300 µl)
Reagent Preparation
Wash Buffer: Dilute 25X Wash Buffer with deionized water in a 1:24 ratio (e.g., 4 ml Wash Buffer + 96 ml deionized water). Store diluted buffer at 2-8°C for up to 3 days.
Assay Procedure
- Bring kit and samples to room temperature.
- Read instructions carefully.
- Prepare the microplate by selecting the necessary wells.
- Add 100 µl Positive Control to two wells.
- Add 100 µl Negative Control to another two wells.
- Add 80 µl Sample Buffer, followed by 20 µl Sample, to each well.
- Incubate the plate at 37°C for 60 minutes.
- Wash wells with Wash Buffer (300 µl per well) five times.
- Add 100 µl Enzyme Conjugate to each well and incubate at 37°C for 45 minutes.
Wash wells again as in step 8. - Add 100 µl TMB Substrate to each well and incubate at room temperature for 10 minutes.
- Stop the reaction with 50 µl Stop Solution per well.
- Read the plate at OD450nm using an ELISA reader.
Result Determination
Criteria for a Valid Assay
Validity: Mean OD of Negative Control - Mean OD of Positive Control ≥ 0.6
Criteria for Positive and Negative Results
Inhibition Rate Calculation
Formula
(Mean OD of Negative Control – Mean OD of Sample) / Mean OD of Negative Control * 100% = Inhibition Rate %
Result Interpretation
- Positive: Blocking Rate ≥ 50%
- Negative: Blocking Rate < 50%
Storage and Expiration
- Storage: Store at 2-8°C, avoid direct sunlight.
- Expiration: 12 months from the date of manufacture.