Echovirus 5: infectious transcripts and complete nucleotide sequence from uncloned cDNA

Echovirus 5 (EV5) may be isolated from various neurological and exanthemati c diseases. To determine the relationship of EV5 to other enteroviruses and for studies of its interactions with the target cell, the complete nucleot ide sequence of EV5 was determined. Three overlapping fragments, collective ly representing the complete genome, were amplified with RT-PCR and sequenc ed. Analysis of the EV5 sequence revealed a typical enterovirus-like organi zation of the genome. To verify that the cDNA generated sequence was derive d from infectious viruses, complete EV5 genomes were amplified in one ampli con by long distance PCR. Transfection of in vitro transcribed RNA from the se amplicons into cell cultures resulted in replicating EV5. Comparison of the overall nucleotide and amino acid sequences demonstrates that EV5 can b e regarded as a coxsackievirus B-like enterovirus. Variable sequences betwe en EV5 and the well characterized coxsackievirus B3 (CVB3) are for the most part observed for amino acid residues that correspond to exposed sequences in the CVB3 capsid. This observation indicates that the reported EV5 strai n recently diverged from group B coxsackieviruses. (C) 1999 Elsevier Scienc e B.V. All rights reserved.