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Genomic Analysis of Recombinant Sabin Clinical Isolates
Authors:Eleni Paximadi  Ioannis Karakasiliotis  Zissis Mamuris  Constantinos Stathopoulos  Vassilis Krikelis  Panayotis Markoulatos
Institution:(1) Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Ploutonos 26 and Aiolou, 41221 Larissa, Greece;(2) Virology Laboratory, Hellenic Pasteur Institute, Athens, Greece;(3) School of Animal and Microbial Sciences, University of Reading, Reading, UK;(4) Technological Educational Institute of Larissa, Larissa, Greece
Abstract:Recombination in Poliovirus vaccine strains is a very frequent phenomenon. In this report 23 polio/Sabin strains isolated from healthy vaccinees or from VAPP patients after OPV administration, were investigated in order to identify recombination sites from 2C to 3D regions of the poliovirus genome. RT-PCR, followed by Restriction Fragment Length Polymorphism (RFLP) screening analysis were applied in four distant genomic regions (5′ UTR, VP1, 2C and 3C–3D) in order to detect any putative recombinant. The detected recombinants were sequenced from 2C to the end of the genome (3′ UTR) and the exact recombination sites were determined with computational analysis. Five of the 23 isolated strains were recombinant in one genomic region, two of them in 2C, isolates EP16:S3/S2, EP23:S3/S1, two in 3D isolates EP6:S2/S1, EP12:S2/S1 and one in 3A isolate EP9:S2/Sl. Point mutations were found in strains EP3, EP6, EP9 and EP12. Recombination specific types and sites re-occurrence along with point mutations are discussed concerning the polioviruses evolution.
Keywords:Enterovirus  Sabin  RT-PCR  Recombinant  Mutation  Sequence
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