title: A simple restriction fragment PCR approach for discrimination of humanpathogenic Old World animal Orthopoxvirus species.
abstract: There are reliable polymerase chain reaction assays available for exclusion of Variola virus from other poxviruses. However, the discrimination of humanpathogenic animal Orthopoxviridae is more challenging because of the high genomic conservation. Based on the variability of the A36R gene, we describe a simple 20 min PCR assay followed by a 1 h digest with 3 different restriction enzymes. This assay enables rapid discrimination between Cowpox virus and Monkeypox virus and discrimination of the most prevalent members of the Vaccinia virus and Camelpox virus. The test was orthopoxvirus specificand did not react with parapox (Orf) virus. Moreover, the amplified fragments were also well suited for additional genotyping by direct DNA sequencing.
authors.affiliation: Innsbruck Medical University, Department of Hygiene, Microbiology and Social Medicine, Fritz-Pregl-Strasse 3, A-6020 Innsbruck, Austria. hartwig.huemer@i-med.ac.at
authors.forename: Hartwig P
authors.initials: HP
authors.lastname: Huemer
authors.forename: Bettina
authors.initials: B
authors.lastname: Hönlinger
authors.forename: Reinhard
authors.initials: R
authors.lastname: Höpfl
mesh_terms: D004279:DNA, Viral / Q000235:genetics*; D018155:Orthopoxvirus / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D015678:Viral Structural Proteins / Q000235:genetics
chemical_list: C400287:A36R protein, Vaccinia virus; D004279:DNA, Viral; D015678:Viral Structural Proteins