OPTIMIZATION OF THE HEMAGGLUTINATION INHIBITION ASSAY FOR H5 AVIAN INFLUENZA STRAINS

Abstract

Antibodies generated in response to influenza vaccine or virus exposure can be detected by the Hemagglutination Inhibition Assay (HAT). Avian influenza strains are more difficult to test than seasonal strains. Previous improvements to the assay include use of horse blood instead of turkey blood. The following experiment used a Design of Experiments (DOE) in an attempt to further optimize the assay by analyzing both serum preparation and assay variables. Phase 1 considered some of the conditions for pretreating test serum: heat inactivation, adsorption with red blood cells, and percentage of Receptor Destroying Enzyme (RDE). Phase 2 examined the assay variables of virus incubation time, temperature, and the percentage of bovine serum albumin (BSA) in buffer, virus, and red blood cells. Phase 3 compared the suggested protocol with the current protocol. The suggested modifications were not found to impact the sensitivity of the assay when examined with reference or experimental sera.