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Although a number of techniques for protein fractionation have been available for the past thirty years, protein electrophoresis has only been recently applied to avian diagnostics. Now routinely run in the Avian & Wildlife Laboratory, EPH has become an integral component in well bird/new bird screens as well as an accessory test in chlamydiosis, sarcocystosis, and aspergillosis testing. Please note that due to methodology changes, not all EPH run by different laboratories are equivalent. Take care in comparing results from different labs and use only lab established species specific reference intervals. Also of note, capillary zone electrophoresis (CZE) is now available and provides a greater resolution of fractions.
Basics on Electrophoresis
Plasma is applied to an electrical field on a gel substrate. Proteins migrate on the gel dependent on their chemical charge. After staining, the protein bands are enumerated by laser densitometry. Interpretations based on extensive reference ranges are included in every evaluation.
Sample Collection and Preparation
Heparinized plasma is preferred for this test. For best results, centrifuge the green top tube and separate the plasma to a small microfuge tube for transport to the lab. Mild hemolysis and lipemia are acceptable. Samples with more moderate levels of either will be run at the discretion of the lab.
When to use the test…
Note that electrophoresis provides the only valid method of albumin quantitation in exotics and many mammalian species. As a key negative acute phase protein, this is a key added value of this method.
Approximately 30% of clinically normal birds will have an abnormal EPH pattern. Thus, EPH is an excellent screening test for well/new bird exams. EPH is a strong accessory test for suspect liver and kidney disease cases as well as chlamydophilosis, sarcocystosis, and aspergillosis. If all other tests fail to lead to diagnosis and the bird is clinically sick, EPH might help lead the way to other testing possibilities.
In reptiles and fish/elasmobranch species, electrophoresis also can reflect acute phase responses. While the association of particular changes with specific diseases has not yet been made, electrophoresis will provide assistance in health assessments and prognostic value.
In mammals, assays for specific acute phase proteins often rival protein electrophoresis for sensitivity for the detection of inflammation. That said, EPH does provide accurate quantitation of albumin and gamma globulins.
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What does an abnormal EPH mean in samples from avian species?
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Capillary Zone Electrophoresis
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References