Kinetics of Rift Valley Fever Virus in experimentally infected mice using quantitative real-time RT-PCR.
Rift Valley Fever (RVF) is an important viral zoonosis in Africa affecting animals and humans. Since no protective vaccines or effective treatments are available for human use, accurate and reliable diagnostic methods are essential for surveillance of the disease in order to implement adequate public health actions. To study the kinetics of the RVF Virus (RVFV) infection, a SYBR Green-based quantitative real-time RT-PCR assay was developed. By using primers targeting the S-segment of RVFV, the detection limit of this assay was estimated to 30 RNA templates. Blood and organs of experimentally infected mice were sampled at different time points and RVFV RNA was quantified. High amounts of RVFV RNA were found in blood, brain, and liver samples shortly after infection with a 1-4 days post infection window for viral RNA detection. Mice developed symptoms after the appearance of serum antibodies, indicating that the host response plays an important role in the outcome of the disease. The RVFV quantitative RT-PCR proved to be a valuable diagnostic tool during the first days of infection, before detectable antibody levels and visual symptoms of RVF were observed.
Swedish Defence Research Agency, Department of CBRN Defence and Security, SE-901 82 Umeå, Sweden; Division of Infectious Diseases, Department of Clinical Microbiology Umeå University, SE-901 85 Umeå, Sweden; Division of Virology, Department of Clinical Microbiology Umeå University, SE-901 85 Umeå, Sweden.
Jonas
J
Näslund
Swedish Defence Research Agency, Department of CBRN Defence and Security, SE-901 82 Umeå, Sweden; Division of Infectious Diseases, Department of Clinical Microbiology Umeå University, SE-901 85 Umeå, Sweden; Swedish Institute for Infectious Disease Control, SE-171 82 Solna, Sweden.
Nina
N
Lagerqvist
Swedish Institute for Infectious Disease Control, SE-171 82 Solna, Sweden.
Åke
Å
Lundkvist
Division of Virology, Department of Clinical Microbiology Umeå University, SE-901 85 Umeå, Sweden.
Magnus
M
Evander
Division of Infectious Diseases, Department of Clinical Microbiology Umeå University, SE-901 85 Umeå, Sweden.
Clas
C
Ahlm
Swedish Defence Research Agency, Department of CBRN Defence and Security, SE-901 82 Umeå, Sweden; National Environment Agency, Environmental Health Institute, 11 Biopolis Way, 06-05/08, Helios Block, Singapore 138667, Singapore.
Göran
G
Bucht
D000818:Animals; D001769:Blood / Q000821:virology; D001921:Brain / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D004195:Disease Models, Animal; D007668:Kidney / Q000821:virology; D007700:Kinetics; D008099:Liver / Q000821:virology; D008168:Lung / Q000821:virology; D051379:Mice; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012295:Rift Valley Fever / Q000503:physiopathology*; D012296:Rift Valley fever virus / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology*; D013154:Spleen / Q000821:virology; D013997:Time Factors
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral