Abstract
BackgroundCowpox virus (CPXV), a rodent-borne Orthopoxvirus (OPV) that is indigenous to Eurasia can infect humans, cattle, felidae and other animals. Molecular characterization of CPXVs isolated from different geographic locations is important for the understanding of their biology, geographic distribution, classification and evolution. Our aim was to characterize CPXVs isolated from Fennoscandia on the basis of A-type inclusion (ATI) phenotype, restriction fragment length polymorphism (RFLP) profiles of atip gene fragment amplicon, and phylogenetic tree topology in conjunction with the patristic and genetic distances based on full length DNA sequence of the atip and p4c genes.MethodsATI phenotypes were determined by transmission electron microcopy and RFLP profiles were obtained by restriction enzyme digestion of the atip gene fragment PCR product. A 6.2 kbp region spanning the entire atip and p4c genes of Fennoscandian CPXV isolates was amplified and sequenced. The phylogenetic affinity of Fennoscandian CPXV isolates to OPVs isolated from other geographic regions was determined on the basis of the atip and p4c genes.ResultsFennoscandian CPXV isolates encoded full length atip and p4c genes. They produce wild type V+ ATI except for CPXV-No-H2. CPXVs were resolved into six and seven species clusters based on the phylogeny of the atip and p4c genes respectively. The CPXVs isolated from Fennoscandia were grouped into three distinct clusters that corresponded to isolates from Norway, Sweden and Finland.ConclusionCPXV is a polyphyletic assemblage of six or seven distinct clusters and the current classification in which CPXVs are united as one single species should be re-considered. Our results are of significance to the classification and evolution of OPVs.
Highlights
Cowpox virus (CPXV), a rodent-borne Orthopoxvirus (OPV) that is indigenous to Eurasia can infect humans, cattle, felidae and other animals
Fennoscandian CPXV isolates produce wild type V+ A-type inclusions The production of ATI was one of the first biological properties used to differentiate CPXV from Vaccinia virus (VACV) [31]
Our results show that all the Fennoscandian CPXVs except CPXV-No-H2 produced the wild type V+ ATI in both Vero and A549 cells (Table 1, Figure 1)
Summary
Cowpox virus (CPXV), a rodent-borne Orthopoxvirus (OPV) that is indigenous to Eurasia can infect humans, cattle, felidae and other animals. Our aim was to characterize CPXVs isolated from Fennoscandia on the basis of A-type inclusion (ATI) phenotype, restriction fragment length polymorphism (RFLP) profiles of atip gene fragment amplicon, and phylogenetic tree topology in conjunction with the patristic and genetic distances based on full length DNA sequence of the atip and p4c genes. Members of the Old World OPVs include Variola virus (VARV), the etiologic agent of smallpox; Vaccinia virus (VACV), the vaccine virus used to eradicate smallpox; Cowpox virus (CPXV), a rodent- borne zoonotic OPV that is indigenous to Eurasia; Monkeypox virus (MPXV), a zoonotic OPV that causes smallpox-like diseases in humans; Ectromelia virus (ECTV), the etiologic agent of mousepox (lab mice); Camelpox virus (CMLV) and Taterapox virus (TATV) [2]. OPV species presumed to be endemic to North America include Raccoonpox virus (RCNV), Volepox virus (VPXV), and Skunkpox virus (SKPV) [1,3]
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