Full Genomic Analysis Of Group A Rotavirus Isolated From An Adult With Acute Gastroenteritis

Group A rotaviruses (family Reoviridae) are major pathogens associated with acute gastroenteritis in infants and young children under 5 years of age worldwide. The rotavirus genome consists of 11 segments of dsRNA, encoding six viral structural proteins (VP1 to VP4, VP6, and VP7) and six nonstructural proteins (NSP1 to NSP6). VP7 and VP4 have been widely used in a dual classification system, establishing 24 G genotypes and 33 P genotypes. Recently, an extended group A rotavirus classification and nomenclature was established to encompass all 11 genome segments, defining genotypes for each genome segment. Squence-based studies targeting all the rotavirus gene segments appear more appropriate to generate conclusive data and support studies on rotavirus evolution.In this paper, sequencing of the genomes and phylogenetic analyses of two group A rotaviruses isolated from an adult with acute gastroenteritis were conducted, the details were as follow:Chapter One: A brief review of rotavirus was described. Genome Structure and Organizationvirion, structural protein and non-structureal protein,pathogenesis, serotypes and a new classification system of group A rotavirus were reviewed.Chapter Two:Isolation and full genomic analysis of group A rotavirus strain CC0912-1. The stool specimens of an adult with acute gastroenteritis detected by group A Rotavirus Colloidal Gold immunochromagraphy Diagnostic Kit was rotavirus positive. MAI 04 cells infected with the supernantant of the fecal sample exhibited typical cytopathic effect. The strain CC0912-1 cultured detected by PAGE also showed the typical group A rotavirus characteristics. Genomic dsRNAs were extracted from the cultured CC0912-1 and the 11 genetic segments were amplified by RT-PCR. The complete genome of the CC0912-1 strain was sequenced and the deduced amino acid sequences were determined. Phylogenetic analyses, based on the nt sequence of the entire ORFs of each of the 11 genome segments were conducted. The complete genotype determination of the CC0912-1 strains analyzed in the study possessed the G10-P[15]-I10-R2-C2-M2-Al 1-N2-T6-E2-H3 genotypes.Chapter Three:Genomic analysis of group A rotavirus strain CC0912-2. Electrophoretic migration patterns of genomic RNAs extracted from the supernatant of group A rotavirus positve fecal specimen detected by PAGE showed there was another rotavirus strain CC0912-2. Genomic dsRNAs extracted from the fecal sample were amplified by RT-PCR. Sequencing of vp7, nsp1 and nsp3 displayed CC0912-2 was G2 rotavirus.