title: Recovery of Swedish Equine arteritis viruses from semen by cell culture isolation and RNA transfection.
abstract: Recovery of infectious Equine arteritis virus (EAV) from the semen of persistently infected Swedish stallions was attempted by classical cell culture isolation and by transfection of extracted total RNA. Whereas virus from semen samples stored for several months at -20 degrees C or from extended semen could only be recovered by transfection of extracted RNA, isolation in cell culture was achieved readily with fresh, unextended semen stored at -70 degrees C or directly used after sampling. In parallel, the viruses were examined in the variable region of the large glycoprotein GP5 by nested RT-PCR and direct nucleotide sequencing. The resulting sequences were placed into a large phylogenetic tree from this region, demonstrating that Swedish strains belonged to very diverse phylogenetic groups. This represents the first report of recovery of infectious EAV from archived semen samples by RNA transfection.
authors.affiliation: Department of Virology, National Veterinary Institute, Uppsala, Sweden. christianm@imr.no
authors.forename: Christian
authors.initials: C
authors.lastname: Mittelholzer
authors.forename: Irja
authors.initials: I
authors.lastname: Johansson
authors.forename: Anna-Karin
authors.initials: AK
authors.lastname: Olsson
authors.forename: Merike
authors.initials: M
authors.lastname: Ronéus
authors.forename: Berndt
authors.initials: B
authors.lastname: Klingeborn
authors.forename: Sándor
authors.initials: S
authors.lastname: Belák
mesh_terms: D000818:Animals; D018174:Arterivirus Infections / Q000662:veterinary* / Q000821:virology; D002460:Cell Line; D006224:Cricetinae; D007240:Equartevirus / Q000302:isolation & purification*; D006734:Horse Diseases / Q000821:virology*; D006736:Horses; D008297:Male; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012661:Semen / Q000821:virology*; D017423:Sequence Analysis, RNA; D013548:Sweden / Q000453:epidemiology; D014162:Transfection
chemical_list: D012367:RNA, Viral