SPADA

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

Sensitivity analysis :

• AMDL = 50,000 copies/mL target region of Variola virus • ≥ 500 bp must receive permission from WHO • insertion into another Orthopoxvirus is prohibited

Establish Probability of Detection at AMDL w/in collection buffer ( ≥ 95%) Establish Probability of Detection at AMDL w/in environmental matrix ( ≥ 95%)

• Inclusivity panel – Variola virus : ≥ 2 strain target regions • at least one from each primary clade • encompass differences in target region • Based on bioinformatic analysis

Ensure all inclusivity strains are detected at 2X AMDL in collection buffer Ensure all inclusivity strains are detected at 2X AMDL in environmental matrix Ensure target is detected at 2X AMDL w/in pool of environmental panel organisms (pools of up to 10 organisms at 10X AMDL for each)

Specificity analysis: • Exclusivity panel – near neighbor ( Orthopoxvirus ): • All poxvirus strains listed in the table (one from each major clade) • See AOAC Website for the most updated list Variola Working Group SMPR: tailor panel to assay based on bioinformatics

Species

Strain

Commercial availability

Vaccinia

Elsree

ATCCVR-1549

Cowpox

Brighton

ATCCVR-302

Ectromelia

Moscow

ATCCVR-1374

Monkeypox

V79-I-005

BEI NR-2324

Monkeypox

USA-2003

BEI NR-2500

Raccoonpox

Herman

ATCCVR-838

Skunkpox

ATCC

Volepox

ATCC

Camelpox

BEI

Taterapox

BEI

ParapoxvirusOrf

Vaccine

Colorado Serum Company

• Any additional strains with greater similarity to the assay’s target region(s) than the strains listed above in the table • Based on bioinformatic analysis Ensure all exclusivity strains are NOT detected at 10X AMDL in collection buffer