Development of a multiplex RT-PCR assay for the identification of recombination types at different genomic regions of vaccine-derived polioviruses
Datum
2016Language
en
Schlagwort
Zusammenfassung
Polioviruses (PVs) are the causal agents of acute paralytic poliomyelitis. Since the 1960s, poliomyelitis has been effectively controlled by the use of two vaccines containing all three serotypes of PVs, the inactivated poliovirus vaccine and the live attenuated oral poliovirus vaccine (OPV). Despite the success of OPV in polio eradication programme, a significant disadvantage was revealed: the emergence of vaccine-associated paralytic poliomyelitis (VAPP). VAPP is the result of accumulated mutations and putative recombination events located at the genome of attenuated vaccine Sabin strains. In the present study, ten Sabin isolates derived from OPV vaccinees and environmental samples were studied in order to identify recombination types located from VP1 to 3D genomic regions of virus genome. The experimental procedure that was followed was virus RNA extraction, reverse transcription to convert the virus genome into cDNA, PCR and multiplex-PCR using specific designed primers able to localize and identify each recombination following agarose gel electrophoresis. This multiplex RT-PCR assay allows for the immediate detection and identification of multiple recombination types located at the viral genome of OPV derivatives. After the eradication of wild PVs, the remaining sources of poliovirus infection worldwide would be the OPV derivatives. As a consequence, the immediate detection and molecular characterization of recombinant derivatives are important to avoid epidemics due to the circulation of neurovirulent viral strains. © 2016, Springer Science+Business Media New York.
Collections
Verwandte Dokumente
Anzeige der Dokumente mit ähnlichem Titel, Autor, Urheber und Thema.
-
Detection of negative and positive RNA strand of poliovirus Sabin 1 and echovirus E19 by a stem-loop reverse transcription PCR
Fikatas A., Dimitriou T.G., Kyriakopoulou Z., Moschonas G.D., Amoutzias G.D., Mossialos D., Gartzonika C., Levidiotou-Stefanou S., Markoulatos P. (2017)In this report a strand specific RT-PCR was established for the detection of the replicative negative RNA strand of poliovirus sabin 1 (Sabin1) and Echovirus 19 (E19) strains. The key for the successful conduction of the ... -
Vaccine derived bi- and multi-recombinant Sabin strains
Paximadi, E.; Karakasiliotis, I.; Bolanaki, E.; Krikelis, A.; Markoulatos, P. (2007)A retrospective analysis of five Sabin intertypic recombinant strains, isolated from human feacal specimens during the time period 1978-1985 in Greece, was performed by RT-PCR, Restriction Fragment Length Polymorphism ... -
Detection of unusual mutation within the VP1 region of different re-isolates of poliovirus Sabin vaccine
Dedepsidis, E.; Karakasiliotis, I.; Paximadi, E.; Kyriakopoulou, Z.; Komiotis, D.; Markoulatos, P. (2006)In the present study, a genomic analysis of full VP1 sequence region of 15 clinical re-isolates (14 healthy vaccinees and one bone marrow tumor patient) was conducted, aiming to the identification of mutations and to the ...