Characterization of rotavirus electropherotypes excreted by symptomatic and asymptomatic infants

Abstract

Human rotavirus isolates from 1100 stool samples were analyzed by polyacrylamide gel electrophoresis, and 48 different migration patterns were detected. Heterogeneity in the migration of segment 10 was observed in both long and short electropherotypes in which three long and two short patterns were identified. In spite of these variations all short and long electropherotypes were subgrouped by enzyme immunoassay as subgroups I and II respectively. Mixed infections were detected in 17% of cases and the subgrouping correlated with the corresponding electropherotypes. The same electropherotypes were present in severe, mild and asymptomatic cases and no electropherotype was particularly associated with greater virulence. Furthermore, the electropherotypes isolated from nosocomial asymptomatic cases were the same as those detected from those admitted with severe diarrhea. It seems unlikely that electropherotyping can be used to identify more virulent strains of rotavirus.

Rotavirus detection by dot blot hybridization assay using a non-radioactive synthetic oligodeoxynucleotide probe

“…The [6,14]. Rotavirus infection was also diagnosed by detecting viral antigens using the enzyme-linked immunoassay (EIA) described previously [31].…”

“…Processing of stool samples for dot blot hybridization assays Viral RNA was extracted from stool samples by two consecutive extraction cycles with phenol and chloroform and precipitated with ethanol as previously described [6,14]. To denaturate the RNA, the samples were resuspended in distilled water, boiled for 5 min and quenched on ice for 2 min.…”

“…The detection of rotavirus in hospitalized patients with acute gastroenteritis is important in ensuring use of adequate therapy and also in preventing nosocomial infections [4][5][6][7].…”

“…This latter method may be particularly useful since it has been suggested that the electrophoretic pattern can be used to trace nosocomial infections using the viral electropherotype as a marker to define a particular viral strain [6,14,15].…”

Usage of Silver-Stained Polyacrylamide Gels Electrophoresis (PAGE) for Detection of Rotavirus Infection by Direct Identification of Viral Nucleic Acid

“…The running buffer consisted of Tris glycine pH 8.8. Discontinuous electrophoresis was carried out as described by Laemlli at 30 mA for 3 hrs at room temperature 8 . Finally, the double stranded RNA was visualized by silver staining.…”