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.
Highlights
Classical swine fever (CSF) is one of the most important diseases of pigs and wild boar worldwide
Knowledge on efficiency of virus rescue by RNA transfection directly compared to conventional virus isolation using samples of Classical swine fever virus (CSFV) infected pigs is still missing
To conclude whether RNA transfection can be recommended in addition to conventional virus isolation, the present study focused on direct comparison of both methods using organ samples of experimentally and field infected pigs, respectively
Summary
Classical swine fever (CSF) is one of the most important diseases of pigs and wild boar worldwide. It is caused by Classical swine fever virus (CSFV), a pestivirus belonging to the family Flaviviridae [1]. CSF outbreaks are associated with enormous economic losses, which are a result of the disease itself as well as of implemented control measures including stamping-out strategy and trade restrictions in the European Union (EU) [2, 3]. To confirm a CSF outbreak in the PLOS ONE | DOI:10.1371/journal.pone.0126806. CSFV Recovery by Transfection of RNA from Organ Samples To confirm a CSF outbreak in the PLOS ONE | DOI:10.1371/journal.pone.0126806 May 11, 2015
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