Abstract
The prevalence of North American orthopoxviruses in nature is unknown and may be more difficult to ascertain due to wide spread use of vaccinia virus recombinant vaccines in the wild. A real time PCR assay was developed to allow for highly sensitive and specific detection of North American orthopoxvirus DNA in animal tissues and bodily fluids. This method is based on the amplification of a 156 bp sequence within a myristylated protein, highly conserved within the North American orthopoxviruses but distinct from orthologous genes present in other orthopoxviruses. The analytical sensitivity was 1.1 fg for Volepox virus DNA, 1.99 fg for Skunkpox virus DNA, and 6.4 fg for Raccoonpox virus DNA with a 95% confidence interval. Our assay did not cross-react with other orthopoxviruses or ten diverse representatives of the Chordopoxvirinae subfamily. This new assay showed more sensitivity than tissue culture tests, and was capable of differentiating North American orthopoxviruses from other members of Orthopoxvirus. Thus, our assay is a promising tool for highly sensitive and specific detection of North American orthopoxviruses in the United States and abroad.
Highlights
The family Poxviridae is divided in two subfamilies: Entomopoxvirinae, which infect insects, and Chordopoxvirinae, which infect vertebrates[1]
The genus Orthopoxvirus (OPXV) is the most relevant in terms of human public health concerns and includes viruses that have been associated with severe febrile, rash illness in humans
The NA OPXVs cluster into their own phylogenetic subclade, as shown in the alignment of the myristylated protein ortholog (Figure 2)
Summary
The family Poxviridae is divided in two subfamilies: Entomopoxvirinae, which infect insects, and Chordopoxvirinae, which infect vertebrates[1]. Genera of the subfamily Chordopoxvirinae that may cause human infections include Orthopoxvirus, Parapoxvirus, Molluscipoxvirus and Yatapoxvirus[1,2]. The genus Orthopoxvirus (OPXV) is the most relevant in terms of human public health concerns and includes viruses that have been associated with severe febrile, rash illness in humans. Infection with vaccinia or cowpox viruses usually only results in localized rash illness. The other viruses within this genus are not currently known to cause human disease
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