SPADA

Specificity analysis (cont.): Environmental aerosol matrix samples: • Method developers should obtain environmental matrix samples that are representative/consistent with the collection method to be used • Considerations include: • Collection medium • Duration of collection • Diversity of geographical areas to be sampled • Climatic/environmental conditions • Seasonal changes • Ensure sufficient replicates to represent environmental condition Ensure aerosol matrix samples do NOT cross-react Environmental panel organisms: • Organisms can be pooled (up to 10 per pool) • Method developer must justify exclusion of specific panel organisms Ensure all organisms are NOT detected at 10X AMDL • If unexpected result, each individual organisms from failed pool must be tested individually at 10X AMDL Variola Working Group SMPR: tailor panel to assay based on bioinformatics

Variola Working Group SMPR: tailor panel to assay based on bioinformatics

Bioinformatic analysis :

• In silico screening on signature sequences • Suggestive of potential performance issues

• Guide necessary additions to wet lab screening panels

• Potential tools for in silico screening: • Simulate_PCR • NCBI tools • FastPCR

Method developer submission should include: Description of sequence databases used in the in silico analysis Description of tool used for bioinformatics evaluation Data confirming selected tool performance based on wet-lab testing • Can be generated retrospectively using published assays List of additional strains to be added to inclusivity or exclusivity panels