Abstract
BackgroundEnterovirus C96 (EV-C96) is a newly named type of enterovirus belonging to species C, and the prototype strain (BAN00–10488) was firstly isolated in 2000 from a stool specimen of a patient with acute flaccid paralysis in Bangladesh. In this study, we report the genomic and phenotypic characteristics of two EV-C96 strains isolated from individuals from the Tibet Autonomous Region of China.MethodsHuman rhabdomyosarcoma (RD), human laryngeal epidermoid carcinoma (HEp-2), and human cervical cancer (Hela) cells were infected with the Tibet EV-C96 strains, and enterovirus RNA in the cell culture was detected with a real time RT-PCR-based enterovirus screening method. The temperature sensitivity of Tibet EV-C96 strains were assayed on a monolayer of RD cells in 24-well plates. Full-length genome sequencing was performed by a ‘primer-walking’ strategy, and the evolutionary history of EV-C96 was studied by maximum likelihood analysis.ResultsStrain 2005-T49 grew in all three kinds of cells, and it was not temperature sensitive. In contrast, none of the three cells produced CPE for strain 2012-94H. Phylogenetic analysis of the two Tibetan viruses, other EV-C96 strains, and EV-C prototypes showed that EV-C96 strains were grouped into three clusters (Cluster1–3) based on their VP1 sequences, which may represent three genotypes. Phylogenetic trees based on the P2 and P3 regions highlighted the difference between Chinese EV-C96 strains and the EV-C96 prototype strain BAN-10488. All Chinese strains formed a cluster separate from BAN-10488, which clustered with CV-A1/CV-A22/CV-A19.ConclusionsThere is genetic variability between EV-C96 strains which suggest that at least few genetic lineages co-exist and there has been some degree of circulation in different geographical regions for some time. Some recombination events must have occurred during EV-C96 evolution as EV-C96 isolates cluster with different EV-C prototype strains in phylogenetic trees in different genomic regions. However, recombination does not seem to have occurred frequently as EV-C96 isolates from different years and locations appear to cluster together in all genomic regions analysed. These findings expand the understanding of the characterization of EV-C96 and are meaningful for the surveillance of the virus.
Highlights
Enterovirus C96 (EV-C96) is a newly named type of enterovirus belonging to species C, and the prototype strain (BAN00–10488) was firstly isolated in 2000 from a stool specimen of a patient with acute flaccid paralysis in Bangladesh
Species Enterovirus A (EV-A)–D correspond to the enteroviruses formerly named Human enterovirus A–D, and species EV-C currently consists of 23 serotypes: three polioviruses (PV) type 1–3, nine group A coxsackieviruses (CV-A1, A11, A13, A17, A19–22, and A24), and 11 new EV-C types, including EV-C95, EV-C96, EV-C99, EV-C102, EV-C104, EV-C105, EV-C109, EV-C113, and EV-C116–C118 [2–7]
Spined for 20 min at 1500 g in a refrigerated centrifuge, and aspirated the supernatant for further use. 0.2 ml stool supernatant was inoculated into human rhabdomyosarcoma (RD), human laryngeal epidermoid carcinoma (HEp-2) and human cervical cancer (Hela) cells and incubated at 36 °C for virus propagation, the cells were examined for the development of EV-like cytopathic effects (CPE) daily, recorded all observations of inoculated and control cultures for at least 7 days [20]
Summary
Enterovirus C96 (EV-C96) is a newly named type of enterovirus belonging to species C, and the prototype strain (BAN00–10488) was firstly isolated in 2000 from a stool specimen of a patient with acute flaccid paralysis in Bangladesh. Enteroviruses (EVs) are small, non-enveloped, singlestranded RNA viruses belonging to the family Picornaviridae, which can be classified to 12 species, including nine enteroviruses, Enterovirus A (EV-A) to EV-H, and EV-J and three rhinoviruses, Rhinovirus A–C [1]. Enterovirus genomes contain approximately 7500 nucleotides consisting of a single open reading frame that is flanked by 5′ and 3′ untranslated regions (5′- and 3′-UTRs). The single open reading frame is translated as a single polypeptide that is autocatalytically cleaved to yield three polyprotein precursors: P1, P2, and P3. Polyprotein P1 is further cleaved to generate capsid proteins VP1–VP4, and P2 and P3 are cleaved to generate non-structural proteins named 2A–2C and 3A–3D, respectively
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
