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.

No abstract

No abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine. Due to genetic variation between the European and the US genotype as well as within both genotypes detection of PRRSV is a diagnostic challenge. This paper reports on a ring test to compare different established reverse transcriptase polymerase chain reaction methods applied routinely in 16 different laboratories in Germany. Three different sets of samples were sent to the laboratories which were to be analysed as follows: (i) basis package: detection of PRRS (yes/no); (ii) differentiation package I: differentiation of EU and US genotypes; and (iii) differentiation package II: differentiation of EU field isolates and EU vaccine strain. A total of 80% of the samples of the basic package were analysed correctly, the analysis of the differentiation package I revealed 61.82% correctly tested samples and the two laboratories that analysed the differentiation package II showed only one correct result. The ring test showed that the majority of incorrect diagnoses were false-negative results.

The aim of this study was to investigate the tissue distribution of two different field isolates and two vaccine strains of porcine parvoviruses (PPV) in infected piglets after transplacental infection. The viral load in 10 different foetal organs was determined by real-time polymerase chain reaction assays with SYBR Green targeting the viral VP2 gene and the genomic c-myc gene in 12 foetuses. The viral load in foetal tissues differed greatly among the different parvoviruses. Between one virulent field isolate compared with the other field isolate and the vaccine strains, the detected viral copy number differed in an order of magnitude of 10(9). The virulent isolate contained PPV in all 10 organs with viral loads varying between 10(11) and 10(15) per 10(6) cells. Concerning the other field isolate and the two vaccine strains, if PPV was detected, in most of the cases the highest viral load was found in foetal kidneys with a maximum viral load of 10(3) per 10(6) cells. Additionally, PPV was found in the heart of one foetus, in the liver and duodenum of one foetus and in the thymus of one foetus with viral loads varying between 10(2.1) and 10(3.5) per 10(6) cells. In completely mummified foetuses with no discriminable organs of foetuses infected with the vaccine strains and the less virulent isolate, PPV was present in very low amounts or even below the detection limit.

No abstract