Molecular characterisation of a rare G8P rotavirus strain detected in an infant with gastroenteritis in Italy
Abstract number: P1887
Medici M.C., Abelli L.A., Martinelli M., Dettori G., Chezzi C.
Objective: The aim of the present study was to perform the molecular characterisation and the phylogenetic analysis of a rare G8P group A rotavirus (GARV) strain detected in Parma, Northern Italy, during a 20042005 rotavirus (RV) epidemiological surveillance.
Methods: Two hundred and seventy three RV-like particles positive stools by electron microscopy (EM) out of 856 stools from children (median age 1 year 8 months, range 1 month 10 years 4 months) hospitalised with gastroenteritis in Parma, Northern Italy, were submitted to PAGE analysis.
Genotyping of 271 GARV strains was carried out on dsRNA extracted from 10% PBS stool suspensions by a nested and/or heminested RT-PCR specific for VP7 and VP4 genes, using pools of G and P type specific primers. One strain (PR/1300/04) displayed G8 specificity and was not typeable in the VP4 gene.
After purification by "Qiaquick Gel Extraction Kit" (QIAGEN, Italy), the VP4, VP6, and VP7 first amplicons of the G8 GARV strain were subjected to sequence analysis with automated sequencer 3730 DNA Analyzer (Applied Biosystems, USA). Phylogenetic analysis was conducted using MEGA version 2.1.
Results: The PR/1300/04 strain exhibited P specificity and long e-type. By sequence analysis, the PR/1300/04 VP7 gene displayed high nucleotide (97.9%) and amino acid (99.4%) identities to bovine Japanese and South African strains, while the VP4 gene was closely related (96% and 99.1% nucleotide and amino acid identity) to the human Italian PA169 strain. The VP6 gene was found to contain subgroup I specificity.
Phylogenetic analysis demonstrated the animal origin of PR/1300/04 VP7 gene and the human origin of the VP4 gene.
Conclusion: The data obtained seem to suggest that the Italian PR/1300/04 strain could be the result of a reassortment between a PA169-like strain with P specificity, long e-type and subgroup I, already circulating in Italy, and a G8 animal strain. The increasing number of reports of atypical RV strains in humans suggests that interspecies transmission of gene segments greatly contributes to the RV genetic evolution.
|Session name:||18th European Congress of Clinical Microbiology and Infectious Diseases|
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