Real-time PCR detection of Parapoxvirus DNA

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 Light-Cycler 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 seal-poxvirus. 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. (c) 2006 American Association for Clinical Chemistry