Enteroviruses (EV) have traditionally been classified by antigenic typing using serum neutralization assay. The latter is time-consuming and labour-intensive and requires a large number of antisera to identify all serotypes. The VP1 gene has been shown to correlate with enteroviruses serotype. So, the virus can be identified by comparison of a partial VP1 sequence of the unknown to that of the database prototype. Generic RT-PCR primers (292/222) have been developed to amplify all human enteroviruses. RT-PCR amplification of the VP1 gene and amplicon sequencing have been used to discriminate among the prototype strains of all human EV serotypes, to identify enteroviruses isolated from human clinical specimens, those were refractory to antigenic typing and to identify potential new untypeable isolates. Infants (6M-5Y) stool samples were collected from different governorates for isolation of enteroviruses. RNA was extracted and used in RT-PCR, amplification reaction carried out with consensus-degenerate primers (292/222) designed from VP1 region, the primers were designed for broad target specificity and amplified all recognized and proposed EV serotypes. The amplification reaction was carried out and the expected and correct molecular size of the PCR product was 338 bp on 1.5% agarose gel. The product of the expected size was successfully amplified and sequenced allowing identification of the infecting virus. The VP1 sequences derived from the RT-PCR products allowed rapid phylogenetic and molecular epidemiologic analysis of the circulating strains during the EV season and comparison with EV sequences from previous seasons or from different locations around the world. The positive isolates were typed by sequencing, 2 out of 16 (12.5%) isolates were HEV (80 and 97), 1/16 (6.25%) was coxsackie B5 and 13/16 (81.25%) were Echoviruses (2, 3, 7, 8, 11, 13, 14, 19, 21 and 29). Echoviruses type 11 and 19 found to be predominating.
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