@article{mbs:/content/journal/jgv/10.1099/vir.0.81179-0, author = "Oberste, M. Steven and Maher, Kaija and Williams, Alford J. and Dybdahl-Sissoko, Naomi and Brown, Betty A. and Gookin, Michelle S. and Peñaranda, Silvia and Mishrik, Nada and Uddin, Moyez and Pallansch, Mark A.", title = "Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses", journal= "Journal of General Virology", year = "2006", volume = "87", number = "1", pages = "119-128", doi = "https://doi.org/10.1099/vir.0.81179-0", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.81179-0", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3′-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68·3 % of isolates, while the HEV-A and HEV-C primers accounted for 9·7 and 11·3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6·5 %) were amplified by more than one primer set and eight isolates (4·3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3′-non-translated region sequences.", }