Virological analysis
Conjunctival swabs were sent to the laboratory of virology
at the Marseille University Hospital. A real-time PCR
assay detecting human enteroviruses was performed
as described previously [6] and enterovirus RNA was
detected (cycle threshold: n=34). Virus isolation was
attempted using Vero, BGM and MA104 cells and is
still in progress, and molecular typing (nested RT-PCR)
was performed as previously described [7], using the
nucleic acid extract of the initial sample. The nested
PCR allowed amplification of a 327 bp partial sequence
of the VP1 gene. Direct sequencing of the amplicon
provided the definitive identification of coxsackievirus
A24 variant (CV-A24v) via BLAST analysis [8]. The
partial sequence obtained was aligned for comparison
with other homologous CV-A24v virus sequences using
Clustal X [9]. Phylogenetic analysis was performed
using neighbour-joining method (Jukes–Cantor algorithm)
in MEGA 5.0 software [10] and confirmed that
the virus detected is CV-A24v (Figure). The sequence
has been deposited in GenBank under accession number
JX196594**.