SPADA

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Specificity analysis (cont.):
 Environmental aerosol matrix samples:
• Method developers should obtain environmental matrix samples that are
Variola Working Group SMPR:
Tailor Panel to Assay Based on Bioinformatics
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
Bioinformatic analysis:
• In silico screening on signature sequences
• Suggestive of potential performance issues
G id dditi t t l b i l
Variola Working Group SMPR:
Tailor Panel to Assay Based on Bioinformatics
• u e necessary a ons o we a screen ng pane s
• Potential tools for in silico screening:
•
• NCBI tools
 Method developer submission should include:
 Description of sequence databases used in the in silico analysis
 Description of conditions used for in silico analysis
 Stringency of in silico analysis must match bench hybridization conditions
 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