rescue of foot-and-mouth disease viruses that are pathogenic for cattle from preserved viral rna samples救援的牛口蹄疫病毒的致病性病毒rna样本保存.pdfVIP

Rescue of Foot-and-Mouth Disease Viruses That Are Pathogenic for Cattle from Preserved Viral RNA Samples 1 1,2,3 1 1 Graham J. Belsham *, Syed M. Jamal , Kirsten Tjørnehøj , Anette Bøtner 1 National Veterinary Institute, Technical University of Denmark, Lindholm, Kalvehave, Denmark, 2 National Veterinary Laboratory, Islamabad, Pakistan, 3 Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan Abstract Background: Foot and mouth disease is an economically important disease of cloven-hoofed animals including cattle, sheep and pigs. It is caused by a picornavirus, foot-and-mouth disease virus (FMDV), which has a positive sense RNA genome which, when introduced into cells, can initiate virus replication. Principal Findings: A system has been developed to rescue infectious FMDV from RNA preparations generated from clinical samples obtained under experimental conditions and then applied to samples collected in the ‘‘field’’. Clinical samples from suspect cases of foot-and-mouth disease (FMD) were obtained from within Pakistan and Afghanistan. The samples were treated to preserve the RNA and then transported to National Veterinary Institute, Lindholm, Denmark. Following RNA extraction, FMDV RNA was quantified by real-time RT-PCR and samples containing significant levels of FMDV RNA were introduced into susceptible cells using electroporation. Progeny viruses were amplified in primary bovine thyroid cells and characterized using antigen ELISA and also by RT-PCR plus sequencing. FMD viruses of three different serotypes and multiple lineages have been successfully rescued from the RNA samples. Two of the rescued viruses (of serotype O and Asia 1) were inoculated