title: Real-time PCR detection of parapoxvirus DNA.
abstract: BACKGROUND
  Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay for the detection of parapoxviruses has been reported.
  
  
  METHODS
  A minor groove binder-based quantitative real-time PCR assay targeting the B2L gene of parapoxviruses was developed on the ABI Prism and the LightCycler platforms.
  
  
  RESULTS
  The real-time PCR assay successfully amplified DNA fragments from a total of 41 parapoxvirus strains and isolates representing the species orf virus, bovine papular stomatitis virus, pseudocowpoxvirus, and sealpoxvirus. Probit analysis gave a limit of detection of 4.7 copies per assay (95% confidence interval, 3.7-6.8 copies per reaction). Scabs contain a sufficient amount of parapoxvirus DNA and can therefore be used for PCR without any DNA preparation step. No cross-reactivity to human, bovine, or sheep genomic DNA or other DNA viruses, including orthopoxviruses, molluscum contagiosum viruses, and yaba-like disease viruses, was observed.
  
  
  CONCLUSIONS
  The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.
authors.affiliation: Robert Koch-Institut, Zentrum für Biologische Sicherheit 1, Berlin, Germany. nitschea@rki.de
authors.forename: Andreas
authors.initials: A
authors.lastname: Nitsche
authors.forename: Mathias
authors.initials: M
authors.lastname: Büttner
authors.forename: Sonja
authors.initials: S
authors.lastname: Wilhelm
authors.forename: Georg
authors.initials: G
authors.lastname: Pauli
authors.forename: Hermann
authors.initials: H
authors.lastname: Meyer
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D018157:Parapoxvirus / Q000235:genetics / Q000302:isolation & purification*; D011213:Poxviridae Infections / Q000662:veterinary / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral