Real-time PCR detection of parapoxvirus DNA

Clin Chem. 2006 Feb;52(2):316-9. doi: 10.1373/clinchem.2005.060335.


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.

Conclusion: The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.

MeSH terms

  • Animals
  • DNA, Viral / analysis*
  • Humans
  • Parapoxvirus / genetics
  • Parapoxvirus / isolation & purification*
  • Poxviridae Infections / veterinary
  • Poxviridae Infections / virology*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sensitivity and Specificity


  • DNA, Viral