IDEXX Laboratories, Inc. Rapid Visual Pregnancy Test Kit

Idexx recommends no more than 30 samples per testing run(4 strips, including controls). Use a timer for all incubation steps.
All reagents must be allowed to come to 18 to 26 °C (64 to 79 °F) before use. Mix reagents by gentle inverting or swirling.

1. Obtain coated plates and record the negative control, positive control and sample positions. If using partial plate, remove only those strips sufficient for samples to be tested. Place the remaining strips, along with the desiccant, in the extra zip lock bag provided and return to 2 to 8 °C (36 to 46 °F).
2. Dispense 100 µL (1 full pipette tip) of ‘negative control’ into the negative control well using the precision pipette.
3. Dispense 100 µL (1 full pipette tip) of ‘positive control’ into the positive control well using the precision pipette.
4. Dispense 100 µL (1 full pipette tip) of sample into the appropriate well using the precision pipette.
5. Dispense 3 drops of reagent 1 (detector solution) into each well.
6. Cover the wells with the plate cover and gently tap the plate 10 times to mix. Start the timer and incubate plate for 7 minutes at 18 to 26 °C (64 to 79 °F).
7. Remove cover and invert plate above a liquid water receptacle (or sink) and flick plate downward with force to discard the liquid from the wells. Make sure that all the liquid is removed from the wells.
8. Wash the plate by filling each well completely with distilled or deionized water. Made sure that the stream of water overflows all wells. Invert the plate and discard the water force. Repeat 2 times, for a total of three washes. Firmly tap residual water from each well onto absorbent material.
9. Dispense 3 drops of Reagent 2 (Conjugate) into each well.
10. Cover the wells with the plate cover and gently tap the plate 10 times to mix. Start the timer and incubate plate for 7 minutes at 18 to 26 °C (64 to 79 °F).
11. Repeat steps 7 and 8.
12. Dispense 3 drops of reagent 3 (TMB Substrate) into each well.
13. Cover the wells with the plate cover and gently tap the plate 10 times to mix.
14. Dispense 3 drops of reagent 4 (Stop Solution) into each well.
15. Gently tap the plate 10 times to mix.
16. Place the plate on a white surface (such as the tray or the carboard sheet provided with the kit or white paper). Take visual read.

In order to have a valid test, the ‘positive control’ well must turn blue in color. If the positive control well does not turn blue, all results for the assay are invalid. Repeat the test following a thorough review of the insert.
The pregnancy status of each animal is determined by comparing the color development of the sample well to the ‘negative control’ well.

• If the color development of the sample well is less than, or the same as, that of the negative control well, the animal is considered not pregnant (open).
• If the color development of the sample well is greater than that of the negative control well, then PAGs are present and the animal is considered pregnant.
• Sample wells which are difficult to interpret visually should be considered doubtful/suspect and pregnancy recheck is recommended.

Pregnant-recheck: Embryonic death is common in early stages of cow pregnancy. PAGs can circulate in cow/heifers for some time after early embryonic death. Pregnancy recheck can be performed by testing another sample taken at a later date or by using another method of pregnancy detection.

Idexx encourages producers to work closely with their veterinarian to develop a reproductive management program appropriate for their operation.