Sequence alignment of target regions for the A36R assay. The alignment of 99 strains was made using BioEdit Sequence Alignment Editor. Those strains within one species that were identical in the primer and probe target regions are condensed; the number in parentheses represents the number of strains included. GenBank sequence accession numbers are as follows (A36R sequences for RPXV-USA64-85A, SKPV-USA1978-WA and VPXV-USA1985-CA are not available in GenBank): Camelpox virus (AY009089, NC_003391), Cowpox virus 1 (HQ407377, HQ420893, HQ420895, HQ420898, X94355, KC813511.1, KC813507.1, KC813506.1, KC813504.1, KC813500.1, KC813498.1, KC813497.1, KC813495.1, KC813493.1, KC813492.1, KC813491.1), Cowpox virus 2 (NC_003663, HQ420894, HQ420896, DQ437593, HQ420897, HQ420899, HQ420900, KC813509.1, KC813502.1, KC813499.1, KC813496.1, KC813494.1), Cowpox virus 3 (KC813512.1, KC813510.1, KC813508.1, KC813505.1, KC813503.1, KC813501.1), Monkeypox virus 1 (AY603973, DQ011156, DQ011157), Monkeypox virus 2 (DQ011155, NC_003310, DQ011154), Vaccinia virus (NC_006998, M35027, AY678276). All other accession numbers can be found in Table S1 in the supplemental material. FOR, forward primer; REV, reverse primer; VARV, variola virus; ECTV, ectromelia virus; TATV, taterapox virus; VACV, vaccinia virus. The probe contains 5′ 6-carboxyfluorescein (FAM).