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Virology Journal
Published in: Virology Journal 1/2011

Open Access 01-12-2011 | Methodology

Detection of North American orthopoxviruses by real time-PCR

Authors: Nadia F Gallardo-Romero, Andres Velasco-Villa, Sonja L Weiss, Ginny L Emerson, Darin S Carroll, Christine M Hughes, Yu Li, Kevin L Karem, Inger K Damon, Victoria A Olson

Published in: Virology Journal | Issue 1/2011

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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.
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Metadata
Title
Detection of North American orthopoxviruses by real time-PCR
Authors
Nadia F Gallardo-Romero
Andres Velasco-Villa
Sonja L Weiss
Ginny L Emerson
Darin S Carroll
Christine M Hughes
Yu Li
Kevin L Karem
Inger K Damon
Victoria A Olson
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2011
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/1743-422X-8-313

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