title: Development of a subgenomic clone system for Kyasanur Forest disease virus.
abstract: Emerging tropical viruses pose an increasing threat to public health because social, economic and environmental factors such as global trade and deforestation allow for their migration into previously unexposed populations and ecological niches. Among such viruses, Kyasanur Forest disease virus (KFDV) deserves particular recognition because it causes hemorrhagic fever. This work describes the completion of an antiviral testing platform (subgenomic system) for KFDV that could be used to quickly and safely screen compounds capable of inhibiting KFDV replication without the requirement for high containment, as the structural genes have been replaced with a luciferase reporter gene precluding the generation of infectious particles. The coordination of KFDV kinetics with the replication characteristics of the subgenomic system has provided additional insight into the timing of flavivirus replication events, as the genetically engineered KFDV genome began replication as early as 2h post cellular entry. Possession of such antiviral testing platforms by public health agencies should accelerate the testing of antiviral drugs against emerging or recently emerged viruses mitigating the effects of their disease and transmission.
authors.affiliation: Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB R3E 3P6, Canada; Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, MB R3E 3L5, Canada; Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada.
authors.forename: Bradley W M
authors.initials: BWM
authors.lastname: Cook
authors.affiliation: Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB R3E 3P6, Canada; Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, MB R3E 3L5, Canada; Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada.
authors.forename: Aidan M
authors.initials: AM
authors.lastname: Nikiforuk
authors.affiliation: Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB R3E 3P6, Canada; Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, MB R3E 3L5, Canada.
authors.forename: Todd A
authors.initials: TA
authors.lastname: Cutts
authors.affiliation: High Containment Respiratory Viruses Group, Special Pathogens Program, National Microbiology Laboratory at the Canadian Science Centre for Human and Animal Health, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3P6, Canada,; Department of Medical Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada.
authors.forename: Darwyn
authors.initials: D
authors.lastname: Kobasa
authors.affiliation: Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada.
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Court
authors.affiliation: Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB R3E 3P6, Canada; Applied Biosafety Research Program, Canadian Science Centre for Human and Animal Health and J. C. Wilt Infectious Diseases Research Centre, Public Health Agency of Canada, 745 Logan Street, Winnipeg, MB R3E 3L5, Canada; Department of Microbiology, The University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada. Electronic address: steven.theriault@phac-aspc.gc.ca.
authors.forename: Steven S
authors.initials: SS
authors.lastname: Theriault
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics* / Q000502:physiology*; D005416:Flavivirus / Q000235:genetics; D017930:Genes, Reporter; D016679:Genome, Viral; D006482:Hemorrhagic Fevers, Viral / Q000175:diagnosis; D057166:High-Throughput Screening Assays; D008156:Luciferases / Q000235:genetics; D012093:Replicon; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D008156:Luciferases
keywords: Antiviral research; Flavivirus; Kyasanur Forest disease virus; Molecular genetics; Subgenomic clone system; Tick-borne flavivirus