ELISA testing for poultry diseases is susceptible to various influences that can affect the accuracy of results, so simply assessing sample results may not provide valid outcomes. To increase precision, the document recommends diluting samples in two steps - first mixing 5 ul of sample with 245 ul of diluent for a 1:50 dilution, then taking 90 ul of that and mixing with 10 ul of the diluted sample for an overall 1:500 dilution. Following correct methods, having good technique and equipment, and using reference controls can help provide more reliable ELISA test results.

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3. •ELISA is susceptible to time, temperature, light,
movement and performance of protocol, so there
are many influencers on the outcome of the test.
Assessing sample results only will not give
guarantee the results are valid.
•Avoid pipetting small volumes: adding 1 ul serum
to 500 ul diluent comes with great risk of being
inaccurate and in precise
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4. •To increase accuracy and precision, it is
recommended to split the sample dilution into
2 steps.
– Step 1: 5 ul sample and add 245 ul diluent
(1:50)
– Step 2: 90 ul diluent and add 10 ul of 1:50
diluted samples (1:10)
•1:50 x 1:10 = 1:500
•By reducing test variation, it allows the
veterinarian to do more meaningful
interpretation of the results
•Get reliable results:
– Correct method
– Good technique
– Good equipment
– Reference control
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