title: Orthopoxvirus pan-genomic DNA assay.
abstract: A genome-spanning assay is described that enables laboratory confirmation of infections with orthopoxviruses (OPVs), particularly Vaccinia, Monkeypox, and Variola viruses, which can cause vesiculo-pustular rash illnesses in humans. The assay uses a series of polymerase chain reaction (PCR) amplicons that overlap to span the approximately 200kilobase pair linear DNA genome of OPVs. Corresponding amplicons of different viral isolates can then be compared by matching their restriction fragment length polymorphism (RFLP) gel electrophoresis patterns. The PCR step does not necessarily require viral growth to produce sufficient DNA for the RFLP comparisons. The assay would be useful as a prelude to sequencing entire or partial DNA genome regions of various OPVs, including natural or recombinant OPVs and potentially dangerous bioengineered OPVs designed to express foreign DNA or other viruses.
authors.affiliation: Poxvirus and Rabies Branch, Division of Viral and Rickettsial Diseases, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Coordinating Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.forename: Susan L
authors.initials: SL
authors.lastname: Ropp
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.forename: Inger K
authors.initials: IK
authors.lastname: Damon
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Esposito
mesh_terms: D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D018155:Orthopoxvirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000592:standards*; D012150:Polymorphism, Restriction Fragment Length; D011213:Poxviridae Infections / Q000175:diagnosis / Q000821:virology; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004279:DNA, Viral