Two Distinct Electrophoretic Migration Patterns of RNA Segments of Human Rotaviruses Prevalent in Japan in Relation to Their Serotypes

Abstract

Polyacrylamide gel electrophoretic analysis of viral nucleic acids has revealed II segments of double-stranded RNA patterns in rotaviruses (8). On the basis of differences in the migration of RNA segments on gels, Espejo et al have demonstrated the presence of two distinct RNA patterns among human rotaviruses obtained from different epidemics (1-5). Most recently, the existence of a correlation between the RNA patterns of human rotaviruses and their serotypes (subgroups) has been demonstrated (6).This communication describes the existence of two distinct RNA patterns among rotaviruses obtained from infants hospitalized with acute diarrhea between 1975 and 1980 in Sendai and Yamagata. Eighteen rotavirus strains were isolated and examined in this study. Extraction and purification of the rotaviruses were carried out by the method developed by Espejo et al (I). As a standard strain, the Wa strain of human rotavirus type 2, which was kindly provided by Dr. R.C. Wyatt (National Institutes of Health, U.S.A.) was used. The Wa strain was grown in MAI04 cells. The purified virus was disrupted at 100 C for 2 min in 0.0625 M Tris-HCl, pH 6.8, containing 1% (w/v) sodium dodecyl sulfate, 4 M urea and 5% (v/v) 2-mercaptoethanol. For electrophoresis of the RNA, 10% (w/v) polyacrylamide-0.27% (w/v) bisacrylamide slab gels were prepared. Appropriate amounts of samples were applied to the gels and electrophoresis was carried out at the constant current of 12 m Ajslab gel for 8 hr. The gels were subsequently stained for 15 min with ethidium bromide solution (I !,g/ml in 0.02 M Tris-HCl, pH 7.4) and photographed. Figure 1 shows the RNA gel electrophoretic profiles of rotavirus strains obtained in Sendai (80S1, 80SR002; 80S2, 80SR004) and in Yamagata (80Yl, 80Y003; 80Y2, 80YOOI) in 1980 and the Wa strain. When compared with the RNA pattern of the Wa strain, the most striking difference in mobility was found in RNA segments 10 and II of the 80S1 and 80S2 strains, which moved significantly more slowly than those of the Wa strain. In contrast, as far as RNA segments 10 and 11 are concerned, no difference was found between the Wa strain and 80Yl or 80Y2. The RNA pattern with slower moving 10th and 11th seg-271

Molecular epidemiology of human rotaviruses: Analysis of outbreaks of acute gastroenteritis in Glasgow and the west of Scotland 1981/82 and 1982/83

“…It was established serologically that viruses of electropherotype L were of subgroup ii, whereas those of the S electropherotype were of subgroup i. This confirms the findings of the literature Kutsuzawa et al 1982;Greenberg et al 1983). All subgroup ii rotaviruses were of serotype 1, and all subgroup i rotaviruses of serotype 2, the pattern most often found in the U.K. in recent years (Sanders, Beards & Flewett, 1983).…”

“…Furthermore, attempts were made to correlate RNA migration patterns with differences observed by serological means Greenberg et al 1981;Kutsuzawa et al 1982;Greenberg et al 1983). This turned out to be difficult, as viruses of the same subgroup or serotype were found to be genomically heterogeneous and, on the other hand, strains of different serotypes within one subgroup showed an indistinguishable RNA migration pattern (Beards, 1982).…”

Rotavirus Infection and Intussusception: A View from Retrospect

“…One final clue is to speculate serotypes from the RNA migration patterns (electropherotypes) of the rotavirus genomes on polyacrylamide gels. Electropherotyping of the rotavirus genome was a cutting-edge technology in the 70s (24), and fortunately I was involved in this type of work (38). Available pictures that contain electropherotypes of the rotaviruses collected at Saiseikan Hospital in the 1974-75 season and 1975-76 season strongly suggest that the circulating strains were serotype G3 (Fig.…”

Genotypic Selection Following Coinfection of Cultured Cells with Subgroup 1 and Subgroup 2 Human Rotaviruses

“…Serotype 2 human strains have been shown to belong exclusively to subgroup 1 and human serotypes 1, 3 and 4 have been found to belong only to subgroup 2 Rodger et al, 1981 ;Hoshino et al, 1984;Svensson et al, 1986;Brown et al, 1988;Georges-Courbot et al, 1988). Subgroup 1 (serotype 2) human rotaviruses have characteristic 'short' electrophoretic patterns associated with an inversion in the migration order of segments 10 and 11 (Kalica et al, 1981a;Kutsuzawa et al, 1982). Several natural isolates have, however, been found to be exceptions (Nakagomi et al, 1987;Brown et al, 1988;Steele & Alexander, 1988;Sethi et al, 1988).…”