title: Classical swine fever virus is genetically stable in vitro and in vivo.
abstract: Phylogenetic analyses of large numbers of classical swine fever strains have revealed a high degree of sequence conservation in the genomic regions examined, suggesting either a recent common ancestor or a low evolution rate. This low variability is in contrast to findings with other RNA viruses. To investigate the consequence of this apparent genetic stability on phylogenetic examinations, the Belgian field isolate Wingene'93 was passaged in pigs as well as in cell culture by various methods. Sequence analyses of viruses collected after various passages in three target regions proposed for phylogenetic studies (5' NTR, E2, and NS5B) revealed a complete sequence conservation. Only when the amount of passaged virus was lowered, mimicking a genetic bottleneck, a single point mutation was observed in the E2 gene. Additionally, only four nucleotide substitutions were observed when the genome of a virus obtained after 96 cell passages in persistently infected cells was compared with its parental virus, the recombinant virus derived from an infectious cDNA clone of CSFV strain Alfort/187. This low mutation frequency observed both in vitro and in vivo demonstrates that classical swine fever virus is genetically stable. Hence, even minor mutations can be considered significant in molecular epidemiological studies.
authors.affiliation: Centrum voor Onderzoek in Diergeneeskunde en Agrochemie (CODA/CERVA), Brussels, Belgium.
authors.forename: H
authors.initials: H
authors.lastname: Vanderhallen
authors.forename: C
authors.initials: C
authors.lastname: Mittelholzer
authors.forename: M A
authors.initials: MA
authors.lastname: Hofmann
authors.forename: F
authors.initials: F
authors.lastname: Koenen
authors.forename: C
authors.initials: C
authors.lastname: Mittelhozer
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006691:Classical Swine Fever / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D014644:Genetic Variation; D016679:Genome, Viral; D010802:Phylogeny; D012367:RNA, Viral / Q000097:blood / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013552:Swine
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral