Genetic Variation in VP7 Gene of Human Rotavirus Serotype 2 (G2 Type) Isolated in Japan, China, and Pakistan

Liu, Wen; Ushijima, Hiroshi; Kakizawa, Junko; Fang, Zhao-Yin; Nishio, Osamu; Morikawa, Shigeru; Motohiro, T

doi:10.1111/j.1348-0421.1995.tb03277.x

Cited by 27 publications

(24 citation statements)

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“…However, substantial differences were observed in the VP8‫ء‬ sequences of two additional P [6] strains, namely, AU19, a unique human G1 rotavirus displaying a supershort RNA electropherotype that was isolated from a child affected with severe diarrhea and dehydration in Japan (26), and the porcine G4 strain, Gottfried, that was originally isolated from the intestinal content of a suckling pig with diarrhea (3). These findings indicate that strains AU19 and Gottfried may be considered prototypes of two additional P [6] lineages (26) and also suggest that the P[6] VP4 gene has undergone genetic and antigenic drift, similar to what has been described for the P[8] VP4 gene and for the G1, G3, G4, G6, and G9 VP7 genes (4,12,16,17,22,24,25,34,36). Interestingly, all three lineages of genotype P [6] represent serologically distinct subtypes, designated P2A [6] (M37-like) and P2C [6] (AU19-like) for the human strains and P2B [6] (Gottfried-like) for the porcine strain (20,26).…”

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Sequencing and Phylogenetic Analysis of Human Genotype P[6] Rotavirus Strains Detected in Hungary Provides Evidence for Genetic Heterogeneity within the P[6] VP4 Gene

et al. 2004

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Although rotavirus genotype P[6] is one of the three most common VP4 specificities associated with human infection, the relatively few sequence data available in public databases suggest that the genetic variability within P[6] might be presently unexplored. Thus far, two human P[6] lineages (M37-like and AU19-like) and a single porcine P[6] lineage (Gottfried-like) have been identified by phylogenetic analysis. Serologic studies demonstrated that these three lineages are antigenically distinct from each other, a finding based on which they were classified into three subtypes, P2A[6] (M37-like), P2B[6] (Gottfried-like), and P2C[6] (AU19-like). To study heterogeneity within this genotype, we selected for molecular characterization a total of six P[6] strains detected during an ongoing surveillance in Hungary. The variable region of the VP4 gene was subjected to sequencing and phylogenetic analysis. Our data indicated that these six strains fell into two phylogenetic lineages distinguishable from the human lineages M37-like and AU19-like and from the porcine lineage Gottfried-like. Further studies are needed to understand whether these two novel lineages are genuine human strains or might have originated from animal strains and to evaluate the antigenic relationship of the novel Hungarian P[6] strains to the three established subtypes.Group A rotaviruses, members of the family Reoviridae, are the major cause of acute dehydrating gastroenteritis in children and young animals (7). The viral genome composed of 11 segments of double-stranded RNA is enclosed in a nonenveloped, icosahedral, triple-layered capsid. The outer capsid antigens, VP7 and VP4, carry the serotype and the genotype specificities and serve as a basis for the dual nomenclature. The specificities of VP7 are designated with a G (VP7 is a glycoprotein), while the specificities of VP4 are designated with a P (VP4 is a protease-sensitive protein). Based on nucleotide and deduced amino acid sequences, at least 15 G genotypes and 23 P genotypes have been established. Due to the lack of adequate immunological reagents (i.e., type-specific monoclonal antibodies or hyperimmune sera), some of the P genotypes have not been characterized serologically and a dual classification by P serotypes (designated with unbracketed numbers) and P genotypes (designated with bracketed numbers) is presently used for VP4 characterization. Out of 23 different P genotypes, only 14 P serotypes and 3 subtypes have been identified (7,14,21,23).Thus far, at least 10 G types (G1 to G6, G8 to G10, and G12) and 11 P types (P[1], P[3] to P[6], P[8] to P[11], P[14], and P[19]) have been identified in humans. Most of these specificities can also be found in animals, suggesting that certain human G and P types, especially those that are rarely detected, might have originated from animal strains either by direct interspecies transmission or by reassortment of cognate genes between heterologous and homologous strains. Serotypes G6 and G8, identified frequently in ruminants, or serotype G5, co...

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Sequencing and Phylogenetic Analysis of Human Genotype P[6] Rotavirus Strains Detected in Hungary Provides Evidence for Genetic Heterogeneity within the P[6] VP4 Gene

et al. 2004

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“…The sensitivity of the first PCR with common serotype primers was about tenfold that of enzyme immunoassay with monoclonal antibody (16), and sensitivity of the second PCR was suspected to be higher. Moreover, we have aggressively continued molecular epidemiological studies on each serotype of group A rotavirus (20,21,25) and want to determine the molecular diversities in each serotype of astrovirus by using RT-PCR and sequencing as well.…”

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confidence: 99%

Determination of Serotypes of Astroviruses by Reverse Transcription‐Polymerase Chain Reaction and Homologies of the Types by the Sequencing of Japanese Isolates

Matsui

Ushijima

Hachiya

et al. 1998

Microbiology and Immunology

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Human standard astroviruses, serotypes 1 to 7, and 35 Japanese isolates were typed by reverse transcription and polymerase chain reaction (RT-PCR) with serotype-specific primers for the first time. The results were identical with those obtained by enzyme immunoassay with serotype-specific polyclonal antibodies, a method which has already been reported. RT-PCR with serotype-specific primers is useful for epidemiological studies of astroviruses where serotype-specific polyclonal antibodies are not available. Two parts of the capsid region, N terminus and C terminus, were sequenced. Serotypes differed in those regions. The N terminus differed less than the C terminus between serotypes. Both the N terminus and C terminus were similar intraserotypically with the exception of serotype-4 isolates which could be divided into A and B subgroups on the basis of their C terminus sequences, which were not known previously. Human astroviruses are non-enveloped, single-stranded RNA viruses which were first identified in 1975 by the electron microscopy of stool specimens from children with diarrhea (1, 8). Not only sporadic cases but also outbreaks of astroviruses have been reported (9,13,19). Astroviruses can be cultured in LLC-MK2 cells and CaCo-2 cells (5,22). From the cultured viruses, seven human astrovirus serotypes have been identified serologically by using serotype-specific polyclonal antibodies (6). Serotype 8 was recently reported in a database (Accession No. Z66541).The complete genomic sequences of human astrovirus 1 were determined in 1994 (7, 23). After that, the total capsid sequences of serotypes 2, 4 and 6 were reported (3, 6, 11,24). The partial sequence of the capsid region of serotype 3 is also known (12). These results enabled us to detect and sequence astroviruses by reverse transcription and polymerase chain reaction (RT-PCR) (16).We obtained and cultured the standard astroviruses, serotypes 1 to 7. We then analyzed the full length of the capsid regions by RT-PCR and sequencing (preparing for submission). From these results, we designed serotypespecific oligonucleotide primers in the capsid region. We then compared the serotypes as determined by RT-PCR and enzyme immunoassay (EIA) using serotypespecific polyclonal antibodies on Japanese strains of astroviruses. Moreover, we compared the nucleotides and the predicted amino acid sequence homologies in two parts of the capsid region.Our main purpose was to verify that RT-PCR with serotype-specific primers is applicable to the epidemiological study of astrovirus serotypes as compared to serotyping by EIA and sequence analysis because serotype-specific antibodies are commercially unavailable and there are limited epidemiological studies. In addition, the sequence of the capsid region is analyzed in two parts of the relatively conservative portion, N terminus, and relatively variable portion, C terminus, to deter-

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“…In addition, we have reported molecular analysis of human G 1, G2 and G3 rotaviruses in Japan, China and Pakistan (20,21,23). In our recent study, G3 rotaviruses were considered to be intraserotypically more heterogeneous than G I and G2 viruses.…”

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Molecular Characterization in the VP7, VP4 and NSP4 Genes of Human Rotavirus Serotype 4 (G4) Isolated in Japan and Kenya

Kudo

Zhou

Cao

et al. 2001

Microbiology and Immunology

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The VP7, VP4 and NSP4 genes of human rotavirus serotype 4 (G4) were analyzed to investigate intraserotypic variations. The techniques used included reverse transcription polymerase chain reaction with subtype specific primers, restriction fragment length polymorphism analysis and sequence analysis. Twelveisolates (nine from Japan and three from Kenya) and two standard strains (Hoehi, Odelia) were G4A P[8] Wa group NSP4. A standard strain (ST3) was G4A P[6] Wa group NSP4 and a strain (VA70) was G4B P[8] Wa group NSP4. These results show G4 rotaviruses can be divided into three combinations at the moment.Key words: Rotavirus, Serotype G4, VP7 gene, VP4 gene, NSP4 gene Rotavirus is a major etiologic agent that causes diarrhea in infants and children worldwide (3). In developing countries, rotavirus infection, coupled with malnutrition, leads to a severe and fatal outcome (1). At present, the genus Rotaviridae is classified into seven groups (groups A to G), based on group specific antigen. Group A rotavirus is the most prevalent (18,22). The genome of rotavirus comprises of 11 segments of double-stranded RNA, each of which encodes a distinct viral protein. Two outer capsid proteins, VP4 and VP7, are involved in virus neutralization. The neutralization specificities of VP4 and VP7 are designated as P and G serotypes for protease-sensitive protein and glycoprotein, respectively (5). Currently, two overlapping classification systems are used for VP4. P serotypes can be determined by P serotype specific monoclonal antibodies, however VP7 comprises of the greater percentage of the outer capsid protein and as such hyper-immune antisera raised to intact viral particles primarily recognize VP7 rather than VP4. For this reason, P typing is usually performed using reverse transcription polymerase chain reaction

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Genetic Variation in VP7 Gene of Human Rotavirus Serotype 2 (G2 Type) Isolated in Japan, China, and Pakistan

Cited by 27 publications

References 18 publications

Sequencing and Phylogenetic Analysis of Human Genotype P[6] Rotavirus Strains Detected in Hungary Provides Evidence for Genetic Heterogeneity within the P[6] VP4 Gene

Sequencing and Phylogenetic Analysis of Human Genotype P[6] Rotavirus Strains Detected in Hungary Provides Evidence for Genetic Heterogeneity within the P[6] VP4 Gene

Determination of Serotypes of Astroviruses by Reverse Transcription‐Polymerase Chain Reaction and Homologies of the Types by the Sequencing of Japanese Isolates

Molecular Characterization in the VP7, VP4 and NSP4 Genes of Human Rotavirus Serotype 4 (G4) Isolated in Japan and Kenya

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