Molecular characterization of some human enteroviruses / AbuTaleb Hassanein Hussein Elsayed ; Supervised Mohamed Abdelhameed Shalaby , Hussein Aly Hussein , Aly Fahmy Mohamed

Thesis (Ph.D.) - Cairo University - Faculty of Veterinary Medicine - Department of Virology

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 speci{uFB01}city and ampli{uFB01}ed 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 ampli{uFB01}ed and sequenced allowing identi{uFB01}cation of the infecting virus

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