title: Transfection of RNA from organ samples of infected animals represents a highly sensitive method for virus detection and recovery of classical swine fever virus.
abstract: Translation and replication of positive stranded RNA viruses are directly initiated in the cellular cytoplasm after uncoating of the viral genome. Accordingly, infectious virus can be generated by transfection of RNA genomes into susceptible cells. In the present study, efficiency of conventional virus isolation after inoculation of cells with infectious sample material was compared to virus recovery after transfection of total RNA derived from organ samples of pigs infected with Classical swine fever virus (CSFV). Compared to the conventional method of virus isolation applied in three different porcine cell lines used in routine diagnosis of CSF, RNA transfection showed a similar efficiency for virus rescue. For two samples, recovery of infectious virus was only possible by RNA transfection, but not by the classical approach of virus isolation. Therefore, RNA transfection represents a valuable alternative to conventional virus isolation in particular when virus isolation is not possible, sample material is not suitable for virus isolation or when infectious material is not available. To estimate the potential risk of RNA prepared from sample material for infection of pigs, five domestic pigs were oronasally inoculated with RNA that was tested positive for virus rescue after RNA transfection. This exposure did not result in viral infection or clinical disease of the animals. In consequence, shipment of CSFV RNA can be regarded as a safe alternative to transportation of infectious virus and thereby facilitates the exchange of virus isolates among authorized laboratories with appropriate containment facilities.
authors.affiliation: EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology,Department of Infectious Diseases, University of Veterinary Medicine Hannover,Hannover, Germany.
authors.forename: Denise
authors.initials: D
authors.lastname: Meyer
authors.affiliation: EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology,Department of Infectious Diseases, University of Veterinary Medicine Hannover,Hannover, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Schmeiser
authors.affiliation: EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology,Department of Infectious Diseases, University of Veterinary Medicine Hannover,Hannover, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Postel
authors.affiliation: EU and OIE Reference Laboratory for Classical Swine Fever, Institute of Virology,Department of Infectious Diseases, University of Veterinary Medicine Hannover,Hannover, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Becher
mesh_terms: D000818:Animals; D002460:Cell Line; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000235:genetics* / Q000302:isolation & purification*; D007958:Leukocyte Count; D010976:Platelet Count; D012367:RNA, Viral; D013552:Swine; D014162:Transfection
chemical_list: D012367:RNA, Viral