The full coding sequences of two novel human enterovirus (HEV)-C serotypes 105 and 116, sampled in the Republic of the Congo in 2010 and in Russia in 2011, were identified in this study. Enterovirus (EV)-105 was closest to EV-104 in the 5′ NTR and to EV-109 in the coding genome region. It had the same unconventional 5′ NTR as EV-104 and EV-109. The non-cytopathogenic EV-116 was phylogenetically close to coxsackievirus (CV)-A1, CV-A19 and CV-A22, which also cannot be propagated in routinely used cell cultures. There were signs of recombination within this subgroup of HEV-C; however, recombination with conventional HEV-C was restricted, implying partial reproductive isolation. As there is also evidence of different permissive replication systems and distinct genetic properties of these subgroups, they may represent subspecies of the HEV-C species or different stages of speciation.