TY - JOUR
TI - Evaluation of seroneutralization and molecular diagnostic methods for
echovirus identification
AU - Kottaridi, C
AU - Bolanaki, E
AU - Siafakas, N
AU - Markoulatos, P
JO - Diagnostic Microbiology and Infectious Disease
PY - 2005
VL - 53
TODO - 2
SP - 113-119
PB - EXCERPTA MEDICA INC-ELSEVIER SCIENCE INC
SN - 0732-8893
TODO - 10.1016/j.diagmicrobio.2005.06.016
TODO - echoviruses; RT-PCR; VP1 gene sequencing; 5 ‘-UTR RFLP;
seroneutralization
TODO - In this study we compared the identification results of 41 echovirus
clinical isolates using RIVM pools (National Institute for Public Health
and the Environment RIVM, Bilthoven, The Netherlands) and reverse
transcription-polymerase chain reaction (PCR) assays. Primer pair
UG(52)-UC53 amplified a 433-bp segment in the 5’ untranslated region.
Restriction enzyme HpaII was used for subgrouping of our isolates into 2
different genetic clusters. Amplification of 315 bp that is located in
5’ end of VP1 gene as well as of a long genomic fragment (1452 bp)
including the VP1 3’ end, the entire coding sequence of 2A, 2B, and the
5’ moiety of the 2C-coding region was achieved by the application of PCR
protocols with primers 292-222 and EUG2a, 2b, 2c-EUC2, respectively.
Phylogenetic trees were constructed for the 5’ end as well as for the 3’
end of VP1 gene using nucleotide sequences derived from sequencing of
clinical isolates and homologous sequences of all echovirus serotypes.
The phylogenetic grouping pattern of the clinical isolates revealed a
correlation of serotype and genotype either in the 5’ or in the 3’ end
of the VP1 gene that was investigated in the present study claiming that
they can be either used for molecular typing of echoviruses. (c) 2005
Elsevier Inc. All rights reserved.
ER -