In the present report, the results of a comparative study in the
detection of all Echoviruses reference strains as well as of 38 clinical
isolates are presented. Using RT-PCR with already published primer pairs
(UG(52)-UC53, 292-222, 012-011 and EUG2a, 2b, 2c-EUC2) from the 5’UTR,
the VP1 region as well as a long genomic fragment including the VP1 3’
end, the entire coding sequence of 2A, 2B, and the 5’ moiety of the
2C-coding region amplification was effective with all reference and
clinical Echovirus isolates with primer pair UG52-UC53 while with
292-222 and 012-011 were amplified 27/28 reference Echovirus strains and
all clinical isolates. As far as EUG2a,2b,2c-EUC2 is concerned, the
RT-PCR gave a positive result for 26/28 reference Echovirus strains and
34/38 clinical isolates. The sequence analysis of a large part of the
5’UTR has revealed that there is no correlation between 5’UTR identity
and the currently recognized human enterovirus species. It has been
suggested that part of VP1 coding sequence would correlate well with
serotype since a number of important neutralization epitopes, as well as
receptor recognition sequences, lie within the VP1 coding sequence.
Therefore, UG52-UC53 and 292-222 primer pairs seem to be the most
appropriate for Echovirus detection and, moreover, UG52-UC53 is useful
for the classification of enteroviruses into genetic clusters
(sub-groups) while 292-222 for the identification of enteroviruses by
amplicon sequencing. (C) 2004 Elsevier Ltd. All rights reserved
