AOAC SPADA Meeting

Assay design process

1. Align all sequences available in  GenBank  ‐ Whole genome for viruses ‐ Literature search for bacteria and  parasites (ex. housekeeping genes) 2. Identify conserved regions to  focus on for assay design ‐ Easier to identify for bacteria ‐ Can be challenging for viruses

Hazara virus alignment

United States Army Medical Research Institute of Infectious Diseases Biodefense solutions to protect our nation

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Assay design: diagnostic assays 3. Identify potential primer/probe combinations that meet the design  and assay requirements ‐ Incorporates assay components and diversity 4. BLAST analysis of the amplicons to triage potential cross‐reactivity ‐ Sometimes not possible due to pathogen diversity → requires secondary assay  to confirm or rule out

5. Wet lab assay down‐selection 6. Assay optimization and testing

United States Army Medical Research Institute of Infectious Diseases Biodefense solutions to protect our nation

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