title: [Development of PCR methods for detection of EAV infection].
abstract: The goal of this work was the development of suitable (real-time) RT-PCR techniques for fast and sensitive diagnosis of EAV and for molecular-epidemiological characterisation of viral strains, as an alternative to virus isolation. To this purpose two conventional RT-PCR methods and one real-time RT-PCR were adapted to detect the broadest possible spectrum of viral strains. Several dilutions with Bucyrus strain showed a 100-fold higher sensitivity of real-time RT-PCR and heminested RT-PCR compared to simple RT-PCR. Making use of 11 cell culture supernatants of different EAV isolates and 7 semen samples of positive stallions, the suitability of the techniques could be shown. Phylogenetic analysis of sequences of the newly analysed samples compared with known sequences indicated that more EAV-lineages exist than presently described.
authors.affiliation: Institut für Veterinär-Virologie, Vetsuisse-Fakultät, Universität Bern.
authors.lastname: Brunner
authors.affiliation: Institut für Veterinär-Virologie, Vetsuisse-Fakultät, Universität Bern.
authors.lastname: Santschi
authors.affiliation: Institut suisse de médecine équine (ISME), Universität Bern und Agroscope, Avenches.
authors.lastname: Gerber
authors.affiliation: Institut suisse de médecine équine (ISME), Universität Bern und Agroscope, Avenches.
authors.lastname: Burger
authors.affiliation: Institut für Veterinär-Virologie, Vetsuisse-Fakultät, Universität Bern.
authors.lastname: Zanoni
mesh_terms: D000818:Animals; D018174:Arterivirus Infections / Q000175:diagnosis* / Q000662:veterinary / Q000821:virology; D007240:Equartevirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006734:Horse Diseases / Q000175:diagnosis* / Q000821:virology; D006736:Horses; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*
keywords: Diagnostik; Equines Arteritis Virus; Phylogenie; diagnosis; equine arteritis virus; phylogeny; real-time RT-PCR