Summary
This kit use PED virus solid phase antigen to detect specific antibody against PED virus antigen, have high sensitivity, specificity, reproducibility, and is easy to operate quickly and used repeatedly.
Reagents and contents
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1.Coated microtiter strips
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1 plate
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7.Stopping solution
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6ml
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2.Conjugate solution
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12ml
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8.Negative control
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800µl
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3.10× Washing solution
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50ml
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9.Positive control
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800µl
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4.Substrate A solution
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6ml
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10.Serological plates
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1 plate
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5.Substrate B solution
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6ml
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11.Microplate Sealers
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2 pieces
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6.Sample dilution
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50ml
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12.Instruction sheet
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1 copy
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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.
Test procedure
1 Diluting concentrated washing solution
Dilute the concentrated washing solution with distilled water or deionized water at 1:10, e.g. add 270 ml distilled water in 30 ml concentrated washing solution, and mix thoroughly.
2 Sample preparation:
Dilute the serum sample with sample dilution at 1:51, e.g. 4 μl serum + 200 μl sample dilution, mix thoroughly.
3 Adding samples and controls
Leave well A1 as reagent blank. Pipette controls and samples as follows:
100 μl of negative control and positive control respectively, and 100 μl diluted samples each into remaining wells.
Incubation at 37 ℃ for 30 minutes. Discard liquid of the wells and fill all wells with diluted washing solution, incubate for 1 minute and discard. Repeat washing procedure two more times as above. At the end of the washing step, carefully remove remaining fluid by tapping the strips on the tissue paper prior to the next step.
4 Adding conjugate solution
Add 100 μl conjugate solution into all wells and incubate at 37 ℃ for 30 minutes. Discard liquid of the wells and wash 3 times as described in step 3.
5 Adding substrates
Add 50 μl substrate A solution and B solution respectively, incubate at 37 ℃ for 10 minutes.
Add 50 μl stopping solution into all wells. Before reading results with ELISA microtiter plate reader, set the reagent blank well A1 to zero . Measure the absorbance of all wells at 450 nm (reference wavelength: 620 nm).
Results
Judgment by microtiter plate reader
Precondition of validity: absorbance value of negative control ≤ 0.1 and absorbance value of positive control ≥ 0.4
Cut-off value =0.15+ (Absorbance value of negative control), absorbance value of negative control should be regarded as 0.05 if it’s below 0.05.
Interpretation of sample results
Absorbance at 450nm ≥ cut-off: PED antibody is positive
Absorbance at 450nm ＜ cut-off: PED antibody is negative
Storage instructions
Store the kit at 2～8℃.Do not freeze any test kit components.
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. recommand to judge results with instruments to improve comparability of testing.