

@article {1749,
	title = {Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses},
	journal = {J Gen Virol},
	volume = {87},
	number = {Pt 1},
	year = {2006},
	note = {Oberste, M Steven Maher, Kaija Williams, Alford J Dybdahl-Sissoko, Naomi Brown, Betty A Gookin, Michelle S Penaranda, Silvia Mishrik, Nada Uddin, Moyez Pallansch, Mark A eng England 2005/12/20 09:00 J Gen Virol. 2006 Jan;87(Pt 1):119-28.},
	pages = {119-28},
	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{\textquoteright}-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{\textquoteright}-non-translated region sequences.},
	keywords = {3{\textquoteright} Untranslated Regions/*analysis/genetics Amino Acid Sequence Capsid/chemistry Capsid Proteins/*genetics DNA Primers Enterovirus/genetics/*isolation \& purification *Genome, Viral Humans Molecular Sequence Data Reverse Transcriptase Polymerase Chain Reaction/*methods Sensitivity and Specificity},
	issn = {0022-1317 (Print) 0022-1317 (Linking)},
	doi = {10.1099/vir.0.81179-0},
	url = {http://www.ncbi.nlm.nih.gov/pubmed/16361424},
	author = {Oberste, M. S. and Maher, K. and Williams, A. J. and Dybdahl-Sissoko, N. and Brown, B. A. and Gookin, M. S. and Penaranda, S. and Mishrik, N. and Uddin, M. and M A Pallansch}
}
