Genomic analysis of recombinant sabin clinical isolates.
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.
Paximadi
Eleni
E
Department of Biochemistry and Biotechnology, School of Health Sciences, University of Thessaly, Ploutonos, Larissa, Greece.
Karakasiliotis
Ioannis
I
Mamuris
Zissis
Z
Stathopoulos
Constantinos
C
Krikelis
Vassilis
V
Markoulatos
Panayotis
P
D000086042:3C Viral Proteases; D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000235:genetics; D019143:Evolution, Molecular; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D017955:Poliovirus / Q000235:genetics* / Q000302:isolation & purification*; D011055:Poliovirus Vaccine, Oral; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics
D020121:5' Untranslated Regions; D036022:Capsid Proteins; D002352:Carrier Proteins; D011055:Poliovirus Vaccine, Oral; D012367:RNA, Viral; C442540:VP1 protein, Poliovirus; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C090443:polymerase 3Dpol, poliovirus; D012324:RNA-Dependent RNA Polymerase; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases; C076023:2C protein, viral