1. Summary

This kit use Canine rabies virus solid phase antigen to detect specific antibody against Canine rabies virus antigen, have high sensitivity, specificity, reproducibility, and is easy to operate quickly and used repeatedly.

2. Reagents and contents

1.         8× Coated microtiter strips: ready to use

2.         2× Conjugate stock solution: ready to use

3.         2× Conjugate diluent: ready to use

4.         1× PBST powder: ready to use

5.         1× Substrate A solution: ready to use

6.         1× Substrate A solution: ready to use

7.         1× Stopping solution: ready to use

（Warning: Stopping solution irritates eyes and skin. Keep out of the reach of children. Upon contact with the eyes, rinse thoroughly with water and consult a doctor.）

8.         1× Positive control: ready to use

9.         1× Negative control: ready to use

10.     1× Instruction sheet

3. Test procedure

1. Conjugate solution preparation: add one conjugate stock solution into one conjugate diluent , mix thoroughly;

2. Solution preparation: Dissolve each bag PBST powder with 500 ml distilled water,

3. Adding samples and controls: Leave well A1 for reagent blank. Pipette controls and samples as follows:

  Add two drops diluent into each well, then add 5 μl serum sample into each well separately, mix evenly. Synchronously add critical control and negative control each 100 μl into each two wells without diluent and add two drops diluent as the blank. incubate in dark at 37 ℃ for 20 minutes, then discard the liquid of the well, add 1 drop washing solution into each well and fill with distilled water promptly, incubate for 30 seconds, then discard the liquid, continue perform another 4 washing cycles as above, then discard the liquid, flap to dry with the absorbent paper.

4. Adding conjugate solution: Dispense 2 drops into all wells (except blank well) and incubate at 37℃ for 20 minutes. Discard the contents of the wells and wash 5 times as described in step 3.

5. Adding substrates: Dispense 2 drops diluted substrate solution into all wells, incubate at 37℃ for 10 minutes.

6. Dispense one drop stopping solution into all wells to stop reaction.

4. Results

1 Observation with naked eyes before adding stopping solution

Negative: thickness of color is more thin than critical control

Positive: thickness of color is equal to or more thick than critical control .

2  Judgment by microtiter plate reader

A450 (patient) ≥ critical control: positive

A450 (patient) < critical control: negative

5. Notes

1 store at 2-8, make the temperature of the kit return to room temperature before use. screw down the cap after use, don't mix caps between diffirent bottles and don't mix components between diffirent kits

2 fill wells with distilled water when washing, 30 seconds each time, pour out the liquid. prohibit  to use tap water or any other water.

3 recommand to judge results with instruments to improve comparability of testing.