Ranavirus phylogeny and differentiation based on major capsid protein, DNA polymerase and neurofilament triplet H1-like protein genes

• Published in: Diseases of aquatic organisms Vol. 85; no. 2; pp. 81 - 91
• Main Authors: HOLOPAINEN, R, OHLEMEYER, S, SCHÜTZE, H, BERGMANN, S. M, TAPIOVAARA, H
• Format: Journal Article
• Language: English
• Published: Oldendorf Inter-Research 10-06-2009
• Subjects: Biological and medical sciences; Capsid Proteins - genetics; DNA-Directed DNA Polymerase - genetics; Freshwater; Fundamental and applied biological sciences. Psychology; Genes, Viral - genetics; Microbiology; Molecular Sequence Data; Neurofilament Proteins - genetics; Phylogeny; Rana esculenta; Ranavirus; Ranavirus - classification; Ranavirus - enzymology; Ranavirus - genetics; Species Specificity; Systematics; Virology; ISEW, Singapore; Italy; FV3; Amphibia; Zoopathogen; Phylogeny; EHNV; Protein; BIV; Virus; Vertebrata; Iridoviridae; Gene; Fish virus; Pathogenic; DNA; Pisces; Host range; ECV; Specific identification; Ranavirus; Molecular systematics; Capsid; Iridovirus; Amphibian virus
• ISSN: 0177-5103; 1616-1580
• DOI: 10.3354/dao02074

In this study, we developed new methods for differentiation of ranaviruses based on polymerase chain reaction and restriction enzyme analysis of DNA polymerase and neurofilament triplet H1-like (NF-H1) protein gene. Using these methods, we were able to differentiate the 6 known ranaviruses--Bohle iridovirus (BIV), European catfish virus (ECV), epizootic haematopoietic necrosis virus (EHNV), European sheatfish virus (ESV), frog virus 3 (FV3) and Singapore grouper iridovirus (SGIV)--with 3 less characterised virus isolates: short-finned eel ranavirus (SERV), Rana esculenta virus Italy 282/I02 (REV 282/I02) and pike-perch iridovirus (PPIV). Doctor fish virus (DFV) and guppy virus 6 (GV6) were distinguished as a group from the other viruses. In addition, all 11 isolates were analysed and compared based on nucleotide sequences from 3 different genomic regions: major capsid protein (MCP), DNA polymerase and NF-H1. The partial DNA polymerase gene was sequenced from all analysed viruses. The complete sequence of the MCP and a fragment of the NF-H1 gene were obtained from BIV, ECV, EHNV, ESV, FV3, PPIV, REV 282/I02 and SERV. With the exception of GV6, DFV and SGIV, the sequence analyses showed only a few variations within the analysed viruses. The sequence data suggest that PPIV, REV 282/I02 and SERV are new members of the genus Ranavirus. The methods developed in this study provide tools to differentiate between closely related ranaviruses of different host and geographical origin.