August 30, 2016 SPADA Meeting Book

Variola virus Signature (Eroded Specificity): Assay Cross-reacts with Cowpox virus Exclusivity panel Species Strain Name Assay 1 ct for 5 ng DNA Assay 2 ct for 5 ng DNA Vaccinia Copenhagen Negative Negative Vaccinia WR Negative Negative N ti Vaccinia ACAM 2000 ega ve Negative Vaccinia BRZ SERRO Negative Negative Cowpox CPXV‐NOR1995‐MAN Negative Negative Cowpox CPXV GER1980‐EP4 19 Negative Cowpox CPXV GER1991‐3 18 Negative Cowpox CPXV_GER1998_2 17 Negative Cowpox CPXV FIN 2000 Negative Ectromelia ECTV Moscow Negative Negative Monkeypox MPXV RCG 2003 358 Negative Negative Monkeypox MPXV USA 2003 044 Negative Negative Raccoonpox RACV V71‐I‐84 Negative Negative Skunkpox SKPV 1991 Negative Negative Volepox VPXV 2004‐CA‐007 Negative Negative Camelpox CMLV‐78‐I‐2379 17 Negative Taterapox (gerbilpox) TATV‐71‐I‐016 16 Negative Parapoxvirus Orf Vaccine for sheep Negative Negative

What We Have Learned About Variola virus Diagnostic Assay Development/Validation

 Bioinformatic analysis should lead design of validation panels

 Inclusivity panel include all Variola virus strains with differences in assay target region

 Exclusivity panel (near neighbor Orthopoxvirus ) contain viruses with assay target regions most similar to Variola virus

 Exceedingly difficult to construct uniform panels for all assays due to high similarity between Orthopoxviruse s

 Simultaneous identification of multiple Variola virus signatures will increase confidence in initial identification/ verification of the pathogen with real-time PCR