SPADA

Current recommendations: tailor panel to assay based on bioinformatics

Sensitivity analysis: Two low dilutions (100 and 10 fg/ µ L) of DNA

Inclusivity panel – Variola virus : ≥ 2 purified viral stocks (at least one from each primary clade) (encompass differences in target region)

Specificity analysis: One high dilution (100 pg/ µ l) of nucleic acid Exclusivity panel – near neighbor ( Orthopoxvirus ): Eurasian: 1 Ectromelia virus 2 Monkeypox viruses (one from each clade) 1 Camelpox virus 5 Cowpox viruses (one from each predicted clade) 1 Taterapox virus 4 Vaccinia viruses (one from each clade) North American: 1 Raccoonpox virus 1 Skunkpox virus 1 Volepox virus

Current recommendations: tailor panel to assay based on bioinformatics

Specificity analysis (cont.): One high dilution (100 pg/ µ l) of DNA

Exclusivity panel (cont.) – other rash-causing illnesses:

2 Varicella Zoster viruses (J clade and E1 clade) 2 Herpes simplex viruses (type 1 and 2) 2 Rickettsia strains

Chordopoxviruses :

1 Parapoxvirus ( Orf )

Negatives:

13 bacteria strains (skin flora) water