Summary
We report the molecular characterization of rotavirus genotype P[8]G5 strains found in fecal specimens collected in four different regions of Brazil, using digoxigenin (dig)-labeled oligonucleotide probes, sequence analysis, and RNA-RNA hybridization. The closest sequence relationships of the neutralization antigens of these strains were to the VP4 protein of P1A[8]G1 strain KU (93.3% identity in amino acids 11 to 282) and to the VP7 protein of G serotype 5 strain OSU (87.6% identity in amino acids 8 to 232). Based on VP7 sequence differences, we designed dig-probes that allowed us to discriminate porcine OSU-like strains from G5 strains isolated from Brazilian infants. The genetic relationships of two P[8]G5 isolates to other rotavirus genogroups were analyzed by RNA-RNA hybridization with [32P]-GTP probes representative of serotypes P1A[8]G1 (Wa), P[8]G3 (AU17), and P9[7]G5 (OSU). The Brazilian P[8]G5 strains showed sequence homology with genes of Wa-like and OSU-like strains, suggesting that these two strains were naturally occurring reassortants between members of the Wa and porcine rotavirus genogroups. The identification of these strains in diverse geographic areas of Brazil underscores their stability and demonstrates the emergence of clinically important rotavirus diarrhea strains by reassortment.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Ando T, Monroe SS, Gentsch JR, Jin Q, Lewis DC, Glass RI (1995) Detection and differentiation of antigenically distinct small round-structured viruses (Norwalk-like viruses) by reverse transcription-PCR and Southern hybridization. J Clin Microbiol 33: 64–71
Beards G, Desselberger U, Flewett T (1989) Temporal and geographical distributions of human rotavirus serotypes, 1983–1988. J Clin Microbiol 27: 2827–2833
Beards G, Xu L, Ballard A, Desselberger U, McCrae MA (1992) A serotype 10 human rotavirus. J Clin Mirobiol 30: 1432–1435
Bishop R, Unicomb L, Barnes G (1991) Epidemiology of rotavirus serotypes in Melbourne, Australia, from 1973–1989. J Clin Microbiol 29: 862–868
Browning GF, Chalmers RM, Fitzgerald TA, Snodgrass DR (1991) Serological and genomic characterisation of L338, a novel equine group A rotavirus G serotype. J Gen Virol 72: 1059–1064
Chen D, Burns JW, Estes MK, Ramig RF (1989) Phenotypes of rotavirus reassortants depend upon the recipient genetic background. Proc Natl Acad Sci USA 86: 3743–3747
Das BK, Gentsch JR, Cicirello HG, Woods PA, Gupta A, Ramachandra M, Kumar R, Bhan MK, Glass RI (1994) Characterization of rotavirus strains from newborns in New Delhi, India. J Clin Microbiol 32: 1820–1822
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12: 387–395
Dunn SJ, Greenberg HB, Ward RL, Nakagomi O, Burns JW, Vo PT, Pax KA, Das M, Gowda K, Rao CD (1993) Serotype and genotypic characterization of human serotype 10 rotavirus from asymptomatic neonates. J Clin Microbiol 31: 165–169
Estes M (1996) Rotaviruses and their replication, In: Fields B, Knipe DM, Howley PM (eds) Fields Virology Vol 2, 3rd ed. Lippincott-Raven Press, Philadelphia, pp 1625–1655
Estes MK, Cohen J (1989) Rotavirus gene structure and function. Microbiol Rev 53: 410–449
Gentsch JR, Glass RI, Woods P, Gouvea V, Gorziglia M, Flores J, Das BK, Bhan MK (1992) Identification of group A rotavirus gene 4 types by polymerase chain reaction. J Clin Microbiol 30: 1365–1373
Gerna G, Sarasini A, Parea M, Arista S, Miranda P, Brussow H, Hoshino Y, Flores J (1992) Isolation and characterization of two distinct human rotavirus strains with G6 specificity. J Clin Microbiol 30: 9–16
Gerna G, Sears J, Hoshino Y, Steele AD, Nakagomi O, Sarasini A, Flores J (1994) Identification of a new VP4 serotype of human rotavirus. Virology 200: 66–71
Glass RI, Gentsch J, Smith JC (1994) Rotavirus vaccines: success by reassortment? Science 265: 1389–1391
Gorziglia M, Larralde G, Kapikian AZ, Chanock RM (1990) Antigenic relationships among human rotaviruses as determined by outer capsid protein VP4. Proc Natl Acad Sci USA 87: 7155–7159
Gouvea V, de Castro L, do Carmo Timenetsky M, Greenberg H, Santos N (1994) Rotavirus serotype G5 associated with diarrhea in Brazilian children. J Clin Microbiol 32: 1408–1409
Gouvea V, Glass RI, Woods P, Taniguichi K, Clark HF, Forrester B, Fang ZY (1990) Polymerase chain reaction amplification and typing of rotavirus nucleic acids from stool specimens. J Clin Microbiol 28: 276–282
Gouvea V, Santos N, do Carmo Timenetsky M (1994) Identification of bovine and porcine rotavirus G types by PCR. J Clin Microbiol 32: 1338–1340
Hardy ME, Gorziglia M, Woode GN (1993) The outer capsid protein VP4 of equine rotavirus strain H-2 represents a unique VP4 type by amino acid sequence analysis. Virology 193: 492–497
Hoshino Y, Sereno MM, Midthun K, Flores J, Kapikian AZ, Chanock RM (1985) Independent segregation of two antigenic specificities (VP3 and VP7) involved in neutralization of rotavirus infectivity. Proc Natl Acad Sci USA 82: 8701–8704
Isa P, Snodgrass DR (1994) Serological and genomic characterization of equine rotavirus VP4 proteins identifies three different P serotypes. Virology 201: 364–372
Kaga E, Iizuka M, Nakagomi T, Nakagomi O (1994) The distribution of G (VP7) and P (VP4) serotypes among human rotaviruses recovered from Japanese children with diarrhea. Microbiol Immunol 38: 317–330
Kapikian AZ, Chanock RM (1990) Rotaviruses. In: Fields B, Knipe DM, Howley PM (eds) Virology, vol 2, 2nd ed. Raven Press, New York, pp 1353–1404
Leite JPG, Alfieri AA, Woods PA, Glass RI, Gentsch JR (1996) Rotavirus G and P Types circulating in Brazil: characterization by RT-PCR, Probe hybridization and sequence analysis. Arch Virol 141: 2265–2274
Li B, Clark HF, Gouvea V (1993) Nucleotide sequence of the VP4-encoding gene of an unusual human rotavirus (HCR3). Virology 196: 825–830
Nagesha HS, Huang J, Hum CP, Holmes IH (1990) A porcine rotavirus strain with dual serotype specificity. Virology 175: 319–322
Nakagomi O, Nakagomi T (1991) Genetic diversity and similarity among mammalian rotaviruses in relation to interspecies transmission of rotavirus. Arch Virol 120: 43–55
Nakagomi O, Nakagomi T, Akatani K, Ikegami N (1989) Identification of rotavirus genogroups by RNA-RNA hybridization. Mol Cell Probes 3: 251–261
Pereira HG, Azeredo RS, Leite JPG, Candeias JAN, Racz ML, Linhares AC, Gabbay YB, Trabulsi JR (1983) Electrophoretic study of the genome of human rotaviruses from Rio de Janeiro, Sao Paulo and Belem, Brazil. J Hyg 90: 117–125
Pereira HG, Linhares AC, Candeias JAN, Glass RI (1993) National laboratory surveillance of viral agents of gastroenteritis in Brazil. Bull Pan Am Health Organ 27: 224–233
Qian Y, Green KY (1991) Human rotavirus strain 69M has a unique VP4 as determined by amino acid sequence analysis. Virology 182: 407–412
Ramachandran M, Das BK, Vij A, Kumar R, Bhambal SS, Kesari N, Rawat H, Bahl L, Thakur S, Woods PA, Glass RI, Bhan MK, Gentsch JR (1996) Unusual diversity of human rotavirus G and P genotypes in India. J Clin Microbiol 34: 436–439
Smith EM, Estes MK, Graham DY, Gerba CP (1979) A plaque assay for the simian rotavirus SA11. J Gen Virol 43: 513–519
Taniguchi K, Pongsuwanna Y, Choonthanon M, Urasawa S (1990) Nucleotide sequence of the VP7 gene of a bovine rotavirus (strain 61A) with different serotype specificity from serotype 6. Nucleic Acids Res 18: 4613
Taniguchi K, Urasawa T, Kobayashi N, Gorziglia M, Urasawa S (1990) Nucleotide sequence of VP4 and VP7 genes of human rotaviruses with subgroup I specificity and long RNA pattern: implication for new G serotype specificity. J Virol 64: 5640–5644
Timenestsky M do C, Santos N, Gouvea V (1994) Survey of rotavirus G and P types associated with human gastroenteritis in Sao Paulo, Brazil, from 1986 to 1992. J Clin Microbiol 32: 2622–2624
Urasawa S, Urasawa T, Wakasugi F, Kobayashi N, Taniguchi K, Lintag IC, Saniel MC, Goto H (1990) Presumptive seventh serotype of human rotavirus. Arch Virol 113: 279–282
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Alfieri, A.A., Leite, J.P.G., Nakagomi, O. et al. Characterization of human rotavirus genotype P[8]G5 from Brazil by probe-hybridization and sequence. Archives of Virology 141, 2353–2364 (1996). https://doi.org/10.1007/BF01718636
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01718636