Arch Virol (2003) 148: 65–81
DOI 10.1007/s00705-002-0908-1
Phylogenetic analysis of rabbit haemorrhagic disease virus
in France between 1993 and 2000, and the characterisation
of RHDV antigenic variants
G. Le Gall-Recul´e
1
, F. Zwingelstein
1
, S. Laurent
2
, C. de Boiss´eson
3
,
Y. Portejoie
4
, and D. Rasschaert
2
1
Agence Fran¸caise de S´ecurit´e Sanitaire des Aliments (AFSSA),
Laboratoire d’
´
Etudes et de Recherches Avicoles et Porcines, Unit´e de Virologie,
Immunologie, Parasitologie, Aviaires et Cunicoles, Ploufragan, France
2
Institut National de la Recherche Agronomique (INRA), Unit´e 086 BASE,
Laboratoire de Virologie et Barri`ere d’Esp`ece, Nouzilly, France
3
AFSSA, Unit´eG´en´etique Virale et Bios´ecurit´e, Ploufragan, France
4
Laboratoire V´et´erinaire D´epartemental du Maine et Loire, Angers, France
Received June 7, 2002; accepted August 12, 2002
Published online November 4, 2002
c
Springer-Verlag 2002
Summary. The first molecular epidemiological study of Rabbit haemorrhagic
disease virus undertaken in France between 1988 and 1995, identified three
genogroups,twoofwhich(G1,G2)disappearedquickly.We used immunocapture-
RT-PCR and sequencing to analyse 104 new RHDV isolates collected between
1993 and 2000. One isolate was obtained in 2000 from a French overseas territory,
the Reunion Island. The nucleotide sequences of these isolates were aligned with
those of some French RHDV isolates representative of the three genogroups
previously identified, of some reference strains and German and American RHDV
antigenic variants. Despite the low degree of nucleotide sequence variation, three
new genogroups (G4 to G6) were identified with significant bootstrap values.
Two of these genogroups (G4 and G5) were related to the year in which the
RHDV isolates were collected. Genogroup G4 emerged from genogroup G3,
which has now disappeared. Genogroup G5 is a new independent group. The
genogroup G6 contained an isolate collected in mainland France in 1999 and the
isolate collected from the Reunion Island, as well as German andAmerican RHDV
variants. Multiple sequence alignments of the VP60 gene and antigenic analysis
with monoclonal antibodies demonstrated that these French isolates are two new
isolates of the RHDV variant.