TY - JOUR

T1 - Quantitation of ranaviruses in cell culture and tissue samples

A1 - Holopainen,Riikka

A1 - Honkanen,Jarno

A1 - Jensen,Britt Bang

A1 - Ariel,Ellen

A1 - Tapiovaara,Hannele

AU - Holopainen,Riikka

AU - Honkanen,Jarno

AU - Jensen,Britt Bang

AU - Ariel,Ellen

AU - Tapiovaara,Hannele

PB - Elsevier BV

PY - 2011

Y1 - 2011

N2 - A quantitative real-time PCR (qPCR) based on a standard curve was developed for detection and quantitation of ranaviruses. The target gene for the qPCR was viral DNA polymerase (DNApol). All ten ranavirus isolates studied (Epizootic haematopoietic necrosis virus, EHNV; European catfish virus, ECV; European sheatfish virus, ESV; Frog virus 3, FV3; Bohle iridovirus, BIV; Doctor fish virus, DFV; Guppy virus 6, GV6; Pike-perch iridovirus, PPIV; Rana esculenta virus Italy 282/I02, REV282/I02 and Short-finned eel ranavirus, SERV) were detected with the qPCR assay. In addition, two fish cell lines – epithelioma papulosum cyprini (EPC) and bluegill fry (BF-2) – were infected with four of the isolates (EHNV, ECV, FV3 and DFV), and the viral quantity was determined from seven time points during the first three days after infection. The qPCR was also used to determine the viral load in tissue samples from pike (Esox lucius) fry challenged experimentally with EHNV.

AB - A quantitative real-time PCR (qPCR) based on a standard curve was developed for detection and quantitation of ranaviruses. The target gene for the qPCR was viral DNA polymerase (DNApol). All ten ranavirus isolates studied (Epizootic haematopoietic necrosis virus, EHNV; European catfish virus, ECV; European sheatfish virus, ESV; Frog virus 3, FV3; Bohle iridovirus, BIV; Doctor fish virus, DFV; Guppy virus 6, GV6; Pike-perch iridovirus, PPIV; Rana esculenta virus Italy 282/I02, REV282/I02 and Short-finned eel ranavirus, SERV) were detected with the qPCR assay. In addition, two fish cell lines – epithelioma papulosum cyprini (EPC) and bluegill fry (BF-2) – were infected with four of the isolates (EHNV, ECV, FV3 and DFV), and the viral quantity was determined from seven time points during the first three days after infection. The qPCR was also used to determine the viral load in tissue samples from pike (Esox lucius) fry challenged experimentally with EHNV.

KW - Viral load

KW - Ranavirus

KW - DNA polymerase

KW - Quantitative real-time PCR

U2 - 10.1016/j.jviromet.2010.11.004

DO - 10.1016/j.jviromet.2010.11.004

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 1

VL - 171

SP - 225

EP - 233

ER -