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

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

Variola Working Group SMPR:

tailor panel to assay based on bioinformatics