Veterinary Diagnostic Kit
WITNESS™ BVDV detects Bovine Viral Diarrhoea Virus (BVDV) Erns antigen in bovine ear tissue samples.
BVDV Erns antigen is detected in ear notch samples from persistently infected (PI) animals. Sensitized particles bound to the BVDV Erns antigens present in a sample from BVDV infected animals migrate along a membrane. The complex is then captured on a sensitized reaction line where its accumulation causes the formation of a clearly visible pink/red band. A control band on the strip ensures that the test was performed correctly.
Before collecting the ear notch, dip the notching tool in disinfectant and then rinse away residual disinfectant with copious quantities of clean water. Select a clean portion of the animal’s ear and avoid blood vessels to harvest a sample. Refer to the table below to obtain a proper ear notch size.
Large ear notch
Smaller ear notch
(≥ 2 - 3 cm and < 1 cm)
Size requirement Ear notch is at least 1 cm on one side Ear notch is at least 2 - 3 mm in diameter and < 1cm in all dimensions
1. SAMPLE PREPARATION
Large ear notch
Small ear notch
(≥ 2 - 3 cm and < 1 cm)
Extraction Buffet 1.5 ml 0.5 ml Mix by inversion 4 times 4 times incubation time
(at 15 °C - 30 °C)
At least 10 minutes (10 minutes to 1 hour) At least 1 hour (1 hour to 3 days) Extracted Sample Storage
If the extracted ear notch samples will not be tested immediatly they should be stored at 2 °C - 7 °C for up to 48 hours.
For prolonged storage, samples should be kept frozen (< -20 °C).
2. BUFFER DISPENSING
First add the appropriate volume of extraction buffer to the graduated tube as follows:
- 1,5 ml of extraction buffer for a large (≥ 1 cm) ear notch
- 0,5 ml of extraction buffer for a small (≥ 2 - 3 mm and < 1 cm) ear notch
Place the ear notch into the graduated tube containing extraction buffer
- Invert the tube containing the ear notch and buffer 4 times to mix, make sure the ear notch is immersed in the buffer.
- Incubate the ear notch samples within the extraction buffer at 15 °C - 30 °C for the appropriate length of time:
- Incubate the large ear notch samples for at least 10 minutes
- Incubate the small ear notch samples for at least 1 hour
3. TESTING SAMPLES
- Use the disposable pipette to dispense 250 μl of the extracted sample into the test tube provided. (Note: To provide approximately 250 μl of extracted sample, insert the pipette into the extraction solution, squeeze the bulb, draw up the liquid from the extraction tube until the liquid reaches the nearest graduated line and then dispense all the liquid into the test tube.)
- Grasp only the labeled portion of the test strips to remove one strip from the desiccant vial for each sample to be tested.
- Place the test strip directly into the test tube so that the red pad on the test strip is submerged in the extracted sample.
- Incubate the test strip in the extracted sample for 15 minutes.
- Remove the test strip from the test tube.
- Read results immediately.
- After 15 minutes, observe the presence or absence of pink/red bands in the center of the test strip between the two absorption pads.
- The control band appears in the upper end of the test strip (closest to the labeled region), while the sample test results are read in the lower part of the test strip.
- Discard the test strip and test tubes in an appropriate biohazard container in accordance with local regulations.
The test is VALID if a pink/red band develops in the upper Control part of the test strip. The absence of the Control band indicates that the test is invalid and must be repeated.
Interpretation of Results
POSITIVE for BVDV: Two pink/red bands (Control band and Test band) are clearly visible on the test strip (C & T). With WITNESS™ BVDV, as with other testing methods, a positive result may not always indicate the animal is persistently infected; the animal might be acutely infected with BVDV. The same animal should be retested after three weeks to confirm its PI status. If the result remains positive, the animal will be identified as PI animal.
NEGATIVE for BVDV: A single pink/red band is present in the upper part of the test strip (C).
Note: The test is complete and may be read before 15 minutes if both control and test pink/red bands are visible. The presence of only the control band on the strip before 15 minutes does not mean that the test is complete, as a test band may appear more slowly than the control band.
- Do not use this kit or any of its components after the expiration date.
- The vial holding the test strips contains a desiccant and should be kept tightly closed when not in use.
- Use the test immediately after removal from the desiccant vial (within 10 minutes).
- Test strips should only be handled in the upper, labeled region. Avoid contact with the surface of the test strip.
- Use a separate pipette for each sample.
- Hold the pipette vertically when dispensing buffer from the extracted sample.
- Extraction buffer is preserved with sodium azide.
- For animal use only.
- For short term storage (up to 48 hours), ear notches should be stored refrigerated at 2 °C – 7 °C.
- If refrigerated storage is not available, small ear notches can be stored (up to 3 days at 15 °C – 30 °C).
- For long term storage, the ear notch samples should be kept frozen (-20 °C or colder).
- Allow samples to come to 15 °C – 30 °C before testing.
- Kit should be stored at 2 °C - 25 °C. Kit should not be frozen.
- 1 Desiccant vial containing 10 test strips
- 2 Bottles Extraction buffer (10 ml each)
- 10 graduated tubes with caps
- 10 Test tubes
- 10 Disposable pipettes
- 1 Rack with holders for 5 test tubes
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