• Medientyp: E-Artikel
  • Titel: Real-Time PCR Detection of Parapoxvirus DNA,
  • Beteiligte: Nitsche, Andreas; Büttner, Mathias; Wilhelm, Sonja; Pauli, Georg; Meyer, Hermann
  • Erschienen: Oxford University Press (OUP), 2006
  • Erschienen in: Clinical Chemistry
  • Sprache: Englisch
  • DOI: 10.1373/clinchem.2005.060335
  • ISSN: 0009-9147; 1530-8561
  • Schlagwörter: Biochemistry (medical) ; Clinical Biochemistry
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  • Anmerkungen:
  • Beschreibung: <jats:title>Abstract</jats:title> <jats:p>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.</jats:p> <jats:p>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.</jats:p> <jats:p>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.</jats:p> <jats:p>Conclusion: The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.</jats:p>