August 30, 2016 SPADA Meeting Book

Sensitivity analysis : • AMDL = 50,000 copies/mL target region of Variola virus • ≥ 500 bp must receive permission from WHO i ti i t th O th i i hibit d • nser on n o ano er r opoxv rus s pro e  Establish Probability of Detection at AMDL w/in collection buffer (≥ 95%)  Establish Probability of Detection at AMDL w/in aerosol environmental matrix (≥ 95%) • Inclusivity panel – Variola virus : ≥ 2 strain target regions • at least one from each primary clade ( Li, et. al. PNAS (2007) Oct. 2;104(40):15787-15792.) • 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) Variola Working Group SMPR: Tailor Panel to Assay Based on Bioinformatics

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 Vaccinia Cowpox

Strain Elsree

Commercial availability

ATCC VR‐1549 ATCC VR‐302 ATCC VR‐1374 BEI NR‐2324 BEI NR‐2500 ATCC VR‐838

Brighton Moscow V79‐I‐005 USA‐2003

Ectromelia Monkeypox Monkeypox Raccoonpox

Herman

Skunkpox Vo epox Camelpox Taterapox l

ATCC ATCC

BEI BEI

Parapoxvirus Orf

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