A simple and rapid method for isolation of caliciviruses from liver of infected rabbits.
Rabbit Haemorrhagic Disease Virus (RHDV), a member of the Caliciviridae family, is the etiologic agent of Rabbit Haemorrhagic Disease (RHD); this viral disease is highly contagious and kills more than 90% of infected adult rabbits. Research on experimental calicivirus infection uses inocula obtained from livers of rabbits dying from calicivirus infection. This implies that caliciviruses have to be purified from liver homogenates. Current methods to isolate caliciviruses from rabbit livers are time consuming. We propose here a new procedure for fast purification of rabbit caliciviruses from liver homogenates that uses centrifugation through an iodixanol gradient. This method offers in approximately 2 h a sample with a high degree of calicivirus purity, as shown by its biochemical and immunocytochemistry analysis, which is also able to kill adult rabbits from RHD within 48 h of inoculation.
Teixeira
Luzia
L
Departamento de Anatomia, Instituto de Ciências Biomédicas Abel Salazar (ICBAS) and Unidade Multidisciplinar de Investigação Biomédica (UMIB), Universidade do Porto, Largo Prof. Abel Salazar 2, 4099-003 Porto, Portugal. Electronic address: luzia_tx@icbas.up.pt.
Marques
Raquel M
RM
Departamento de Anatomia, Instituto de Ciências Biomédicas Abel Salazar (ICBAS) and Unidade Multidisciplinar de Investigação Biomédica (UMIB), Universidade do Porto, Largo Prof. Abel Salazar 2, 4099-003 Porto, Portugal.
Águas
Artur P
AP
Departamento de Anatomia, Instituto de Ciências Biomédicas Abel Salazar (ICBAS) and Unidade Multidisciplinar de Investigação Biomédica (UMIB), Universidade do Porto, Largo Prof. Abel Salazar 2, 4099-003 Porto, Portugal.
Ferreira
Paula G
PG
Departamento de Anatomia, Instituto de Ciências Biomédicas Abel Salazar (ICBAS) and Unidade Multidisciplinar de Investigação Biomédica (UMIB), Universidade do Porto, Largo Prof. Abel Salazar 2, 4099-003 Porto, Portugal.
D000818:Animals; D002498:Centrifugation / Q000662:veterinary; D017929:Hemorrhagic Disease Virus, Rabbit / Q000302:isolation & purification*; D008099:Liver / Q000821:virology*; D011817:Rabbits; D014283:Triiodobenzoic Acids
D014283:Triiodobenzoic Acids; C044834:iodixanol