Rotavirus G Research Articles

Novel group A rotavirus G8 P[1] as primary cause of an ovine diarrheic syndrome outbreak in weaned lambs

Rotavirus is a worldwide major cause of diarrhea outbreaks in neonatal ruminants. An outbreak of ovine diarrheic syndrome (ODS) in 50–75 days-old lambs (weaned lambs) is described. Fecal immunochromatography and intestinal immunohistochemistry for rotavirus group A were performed. In addition, semi-nested multiplex RT-PCR for G and P rotavirus genotyping in combination with sequencing were performed, to support the diagnosis and identify the viral strain. A novel ovine rotavirus group A G8 P[1] strain was determined as the main cause of the ODS observed, whereas other pathogens were ruled out.

Novel Group a Rotavirus G8 P[1] as Primary Cause of an Ovine Diarrhoeic Syndrome Outbreak in Weaned Lambs

Novel Group a Rotavirus G8 P[1] as Primary Cause of an Ovine Diarrhoeic Syndrome Outbreak in Weaned Lambs

Molecular epidemiology of human astrovirus infections in Saudi Arabia pediatric patients

Earlier work Tayeb et al. [Tayeb et al. (2008): J Med Virol 80: 1919-1929] set out to study the epidemiology of diarrhea viruses in pediatric populations. The study addressed initially rotavirus, enteric adenovirus, and astrovirus but was later expanded to include norovirus (NoV). Viruses were sought in fecal specimens and characterized for genotype using molecular methods (PCR, RT-PCR, and RFLP) for the first time in KSA. The survey focused on three locations; Jeddah, Makkah, and Riyadh. During the Hajj, the chief population fluxes are via Jeddah to Makkah. One thousand samples were obtained from children (aged 6 years or less) presenting with diarrhea and thus representing community acquired rather than nosocomial infections. Rotavirus was identified in 6% of the samples followed by NoV accounted for 3.5%, astrovirus 1.9%, and adenovirus 1.4%. Rotavirus G9 was characterized for the first time in Saudi Arabia. Adenoviruses were confirmed and further typed using hexon-specific PCR and RFLP. These data were published by Tayeb et al. [Tayeb et al. (2008): J Med Virol 80: 1919-1929]. However, the nature of astrovirus identified was not investigated further. Therefore, more analysis details are appropriate for astrovirus in the Kingdom. As an extension of earlier work carried out on astrovirus serotype distribution in the Kingdom [Tayeb et al. (2008): J Med Virol 80: 1919-1929], a major objective of the project is to use molecular methods to determine the distribution of astrovirus genotype. Such data will help to provide valuable insights into genetic identities and possible sources of virus strains involved not only in pediatric gastroenteritis but also possible outbreaks in the community.

Sustained Efficacy of the Pentavalent Rotavirus Vaccine, RV5, up to 3.1 Years Following the Last Dose of Vaccine

Rotavirus gastroenteritis (RVGE) is a common cause of childhood hospitalizations and emergency department (ED) visits. In the Rotavirus Efficacy and Safety Trial (REST), the pentavalent rotavirus vaccine (RV5) significantly reduced RVGE-associated hospitalizations and ED visits for up to 2 years following the last vaccine dose. This study evaluated whether RV5 remained efficacious beyond 2 years. A total of 20,736 infants from Finland, initially in REST, were followed for RVGE-associated hospitalizations and ED visits in a Finnish extension study (FES) for up to 3.1 years after vaccination (age, ∼3.5 years). The FES added >18,500 person-years and captured 150 RVGE-associated hospitalizations and ED visits (11 RV5; 139 placebo). In REST + FES, RV5 reduced RVGE-associated hospitalizations and ED visits, regardless of rotavirus serotype, by 94.0% (95% confidence interval [CI]: 91.4%-95.9%) for up to 3.1 years after vaccination. RV5 also conferred significant protection against hospitalizations and ED visits associated with rotavirus serotypes G1 (95.5%; 95% CI: 92.8%-97.2%), G2 (81.9%; 95% CI: 16.1%-98.0%), G3 (89.0%; 95% CI: 53.3%-98.7%), G4 (83.4%; 95% CI: 51.2%-95.8%), and G9 (94.2%; 95% CI: 62.2%-99.9%). Rate reductions (95% CI) in hospitalizations and ED visits during the first, second, and third years of life were 94.0% (90.0%-96.5%), 94.7% (90.7%-97.2%), and 85.9% (51.6%-97.2%), respectively. RVGE-associated hospitalizations and ED visits remain common in the second year of life but decrease in the third year of life. RV5 showed sustained protective efficacy against RVGE-associated hospitalizations and ED visits, regardless of rotavirus serotype, for up to 3.1 years after vaccination.

Molecular characterization of unusual bovine group A rotavirus G8P[14] strains identified in western India: Emergence of P[14] genotype

A total of 78 fecal specimens were collected from both apparently healthy ( n = 71) and diarrheic ( n = 7) cattle from an organized farm in Pune, western India in December 2007–January 2008. Three specimens tested positive for group A rotavirus (RV) by antigen capture ELISA were subjected to RT-PCR for amplification of entire coding regions of three structural (VP4, VP6 and VP7) and one nonstructural (NSP4) genes. All three strains were genotyped as G8P[14]. Phylogenetic analysis of the VP7 and VP4 genes showed clustering of the VP7 gene with G8 strains of bovine origin and VP4 gene with P[14] strains of human origin. The identification of VP6 and NSP4 genes to have I2 (subgroup I) and E2 (genotype A) specificity, respectively of bovine and human origin indicated independent segregation of genes in bovine RV strains. This study indicates circulation of a rare RV genotype, G8P[14] in western India. To our knowledge, this is the second report on RV G8[14] isolated from bovine species after bovine group A RV strain, SUN9 from Japan.

Characterization of Human Rotavirus Strains from Children with Diarrhea in Nairobi and Kisumu, Kenya, between 2000 and 2002

Rotavirus infection is a major cause of diarrheal illness and hospitalization in children <5 years old in Kenya and has been described in various settings and locations across the country and for different time points. In this study, we expand on the molecular characterization of rotavirus strains collected in Nairobi and Kisumu, Kenya, between 2000 and 2002. Rotavirus strains were typed by reverse-transcription polymerase chain reaction and characterized using VP6 monoclonal antibodies and RNA electrophoresis of the viral genome. A large proportion of specimens could not be genotyped; 41% did not produce a G type result, and 43% did not produce a P type result. Of the strains that could be genotyped, G1P[8] strains were predominant, followed by G2P[4] strains. In addition, G8 and G9 strains were seen in similar proportions Interestingly, the G and P combinations were more diverse among G8 and G9 rotavirus strains, suggesting the recent introduction of these strains into the human population. These observations are a link between the occasional observation of G8 and G9 strains at the turn of the century and the high predominance of G9P[8] strains observed in Kenya in 2005.

Emergence and Characterization of Serotype G9 Rotavirus Strains from Africa

Serotype G9 strains have been detected sporadically and in localized outbreaks in various African countries, including South Africa, Botswana, Malawi, Kenya, Cameroon, Nigeria, Ghana, Guinea-Bissau, Libya, and Mauritius. Serotype G9 strains were analyzed to investigate genogroup characteristics, including subgroup specificity, electropherotype, and P and G genotypes. In addition, the antigenic composition of the South African G9 strains was assessed. African G9 strains were associated with both DS-1-like characteristics and Wa-like characteristics, indicating the predisposition of G9 strains to frequently reassort. Despite these reassortment events, serotype G9 strains appear to maintain antigenic character in the outer capsid protein, as evident with the reaction of the South African G9 strains with the G9-specific monoclonal antibody F45:1. Phylogenetic analysis clustered African G9 strains geographically, regardless of genogroup characteristics, into 1 lineage (IIId). Two groups of G9 strains, originating in India and Japan, were identified in this lineage. Continuous surveillance of circulating rotavirus strains in Africa is vital to prepare for future vaccine implementation on a continent that clearly needs such preventative medicines.

Diversity of Rotavirus Strains Circulating in West Africa from 1996 to 2000

followed by G3 (19.5%), G2 (13.6%), and G9 (10.3%). The common G-P combinations were G1P[6] (11.2%), G2P[6] (9.8%) and G9P[8] (8.0%). Considerable G type fluctuation was detected from year to year in the subregion, as well as country-specific variation in circulating strains over time. Rotavirus G9 strains were detected with increasing frequency across the region, but rotavirus G4 strains were seen only in Cameroon, and strains with P[4] specificities were relatively rare, except in Ghana and Cameroon. Strains carrying unusual G-P combinations, such as G2P[6], G3P[6], and G9P[6], were also common.

Characterization of Human Rotavirus Recovered from Children with Acute Diarrhea in Kinshasa, Democratic Republic of Congo

Between July and October of 2003, 2004, and 2005, outbreaks of acute gastroenteritis occurred among children <5 years of age in Kinshasa, Democratic Republic of the Congo. Stool specimens (67 in 2003, 108 in 2004, and 116 in 2005) were collected and screened for rotaviruses using either latex agglutination (Diarlex LAA; Orion Diagnostics) or enzyme immunoassay (IDEIA; DakoCytomation). The molecular characteristics of the rotavirus strains were then determined. Group A rotavirus was detected in 195 (76%) of 258 stool specimens. Polyacrylamide gel electrophoresis was used to observe the 11 rotavirus double-stranded RNA segments in 83% of the 195 rotavirus-positive specimens. Six rotavirus group A electropherotypic patterns were noted, predominantly within the short classic pattern (111 [69%]) and the long pattern (37 [23%]). Mixed patterns were noted in the 14 remaining specimens (9%). Of the 29 samples subjected to subgrouping VP6 enzyme immunoassay, subgroup I predominated. Some of the specimens collected in 2003 (n = 26), 2004 (n = 38), and 2005 (n = 52) were analyzed by reverse-transcription polymerase chain reaction, which showed that t G8P[6] and G8P[8] strains predominated in 2003, and G1P[6] strains with short electropherotypic patterns predominated in 2004 and 2005. The emergence in Kinshasa of G8 serotypes, unusually associated with the P[6] genotype, as well as uncommon G1 rotavirus strains showing a short RNA pattern, is significant in relation to the introduction of a rotavirus vaccine and underscores the need for continued rotavirus serotype surveillance in the Democratic Republic of the Congo.

Detection and genotypingof rotavirus among children under 5 years old hospitalized with diarrhea in Tianjin

：Objective To study the prevalence and genotypes of rotavirus(RV) among children,< 5 years old hospitalized with viral diarrhea in Tianjin. MethodsStool specimens were collected from hospitalized diarrhea children in Tianjin children'shospital between May 2008 and April 2009. Detection of rotavirus was employed by ColloidalGold Device. The detected positives were inoculated to MA-104 cells. The total RNA ofvirus was extracted after CPE which was caused by rotavirus were observed, The VP7serotypes were determined by using RT-PCR to amplify the VP7 gene and sequencing theRT-PCR products.The clinical data for each patient were also collected. Results Among 837specimens, the RV antigen positive rate was 26. 3% (220/837). Among all the children withrotavirus diarrhea, 90. 5% (199/220)were < 2 years old. The prevalence of rotavirusdiarrhea in children peaked during Oct. 2008 through Apr.2009. Of the 208 rotaviruspositive specimens, 95 were successfully identified by RT-PCR Thirty-five positive strainsof RV were sequenced, and the sequencing results showed that 32 positive strains werebelonged to rotavirus G1 type, 2 positive strains were belonged to rotavirus G3 type and 1positive strain were belonged to rotavirus C9 type. Conclusion RV was the dominantetiological agent for infantile diarrhea infection in Tianjin, and the predominantserotype was G1. Key words: Rotavirus infections; Rotavirus;

Separation of rotavirus double-layered particles and triple-layered particles by capillary zone electrophoresis

Cell culture derived rotavirus preparations contain a mixture of double-layered particles (DLPs) and triple-layered particles (TLPs). Characterization of rotavirus vaccine products is important to demonstrate a consistent manufacturing process. A capillary zone electrophoresis (CZE) method was developed to separate and quantitate rotavirus DLPs and TLPs in cell lysate samples and CsCl-purified vaccine preparations of each of the five reassortant rotavirus vaccine strains (G1, G2, G3, G4 and P1) contained in the pentavalent rotavirus vaccine, RotaTeq®. The CZE electropherograms showed that migration of DLPs and TLPs from both CsCl-purified and cell lysates resulted in a separation distance of approximately 3min between the two rotavirus particle types. The identification of the peak(s) containing TLPs was confirmed for both CsCl-purified and cell lysate samples by treatment of the samples with 50mM EDTA, which converted TLPs to DLPs. The migration pattern of the DLPs was consistent (23–24min) among all reassortant strains tested, whether the DLPs were CsCl-purified or from cell lysates. However, the migration pattern of the TLP electropherograms of the reassortant rotavirus strains in cell lysates differed from those of the CsCl-purified reassortant rotavirus strains. In the cell lysate samples, the TLPs of the G1 and G2 reassortant rotavirus strains migrated slower that the corresponding TLPs from the CsCl-purified samples, while the migration time of the TLPs of the G3, G4 and P1 reassortants strains from the cell lysate and CsCl-purified samples appeared similar. Also, the TLPs from the CsCl-purified samples appeared as a defined single peak, while most of the TLPs from the cell lysate samples appeared as a broad peak or as multiple peaks. All the migration patterns were reproducible and consistent. Taking into account reproducibility, objective quantitation, and minimal sample manipulation as well as volume, CZE allowed consistent and quantitative characterization of rotavirus vaccine preparations, which is required for evaluation of vaccine products, including process validation.

Changing profile of the bovine rotavirus G6 population in the south of Ireland from 2002 to 2009

Bovine group A rotavirus is one of the main causes of neonatal diarrhoea in calves. This study examined the different G and P genotypes circulating in the bovine population, from 2002–2009, in the south of Ireland. Rotavirus positive bovine faecal samples ( n = 332) were collected from the Cork Regional Veterinary Laboratory, between 2002 and 2009 and subjected to RNA extraction, PAGE analysis, and G and P genotyping. Genotyping analysis identified G6, G10, P[5], and P[11] to be the predominant G and P genotypes in the present study, with G6 rotavirus responsible for 70–80% of rotavirus infections. The highest combination of G and P types found was G6 P[5], followed by G6 P[5+11] mixed infection. The prevalence of G6 and G10 has shifted over the years, with an increase in the amount of G10 P[11] being detected. Novel combinations (G6+G10P[11], G6+G10P[5+11] and G10P[5+11]) were also detected for the first time. In addition to this, sequence analysis of the VP7 RT-PCR amplicons has revealed that Irish G6 strains are falling within three different lineages, III–V. During this study, two samples, initially genotyped as G8P[11] were identified through sequence analysis as being true G6, lineage III with a high nucleotide identity to Hun4, a G6 human sample from Hungary. The increase in novel G and P type combinations, as well as changes seen in G6 samples could have an impact on rotavirus vaccination programmes, as the current vaccine available may not offer protection against all of these circulating types.

Sequence analysis of the VP7 gene of human rotaviruses G2 and G4 isolated in Japan, China, Thailand, and Vietnam during 2001–2003

Sequence and phylogenetic analyses of the rotavirus VP7 gene were performed on 52 human G2 and G4 strains isolated in Japan, China, Thailand, and Vietnam during 2001-2003. All genotype G2 strains included in the study clustered into lineage II of the phylogenetic tree, together with the majority of global G2 strains detected since 1995. The amino acid substitution at position 96 from aspartic acid to asparagine was noted among the emerging or re-emerging G2 rotavirus strains in Japan, Thailand, and Vietnam during 2002-2003. Genotype G4 strains detected in Vietnam grouped into lineage Ia of the phylogenetic tree, whereas Japanese G4 strains clustered in lineage Ic which included emerging G4 strains from Argentina, Italy, Paraguay, and Uruguay. It is noteworthy that an insertion of asparagine was found at position 76 in all the Japanese strains and that its presence might be involved in the emergence of G4 rotavirus in Japan during 2002-2003.

Convalescent phase sera from children infected with G12 rotavirus cross-neutralize rotavirus strains belonging to the Wa genogroup

The emergence of G12 rotaviruses raises questions about the ability of candidate vaccines in providing protection against such emerging genotypes. Therefore, we assessed cross-neutralization against four reference rotavirus strains namely Wa (G1P[8]), DS-1 (G2P[4]), 116E (G9P[11]) and RV024 (G12P[6]) using paired sera from 28 children infected with G1P[8], G2P[4], G9P[6/8] or G12P[6] genotypes. Convalescent sera of G12P[6]-infected children demonstrated heterotypic response against 116E and Wa strains (50 and 33.3 %). In contrast, none of the four G2P[4]-infected children seroconverted against Wa or RV024 rotaviruses. The geometric mean neutralizing antibody titre in convalescent sera of G12P[6]-infected children was eightfold higher against strains belonging to the Wa genogroup (i.e. G1, G9 and G12 rotavirus) than against strains belonging to the DS-1 genogroup (G2 rotavirus). In conclusion, this study demonstrates that neutralization in part may be genogroup specific, and thus a monovalent vaccine based on the Wa genogroup is likely to protect against the G12 rotaviruses.

Molecular characterization of VP1-3 and NSP1-3 genes of porcine group A rotavirus G12 strain RU172: Evidence for porcine origin of human G12 strains

Molecular characterization of VP1-3 and NSP1-3 genes of porcine group A rotavirus G12 strain RU172: Evidence for porcine origin of human G12 strains

Identification of two sublineages of genotype G2 rotavirus among diarrheic children in Parauapebas, Southern Pará State, Brazil

On a world scale, group A human rotaviruses are the most common cause of severe acute gastroenteritis during infancy and childhood, including five (G1, G2, G3, G4, and G9) epidemiologically important genotypes. Among these, G2 denotes a different genogroup which appears to have a cyclic pattern of occurrence and yet little information is available about its genetic variability. The aim of this report was to characterize the emergence of G2 genotype in Paraupebas, Southern Pará State, Brazil, some of which detected after introduction of rotavirus vaccine. A total of 241 fecal specimens from young children with acute gastroenteritis were collected from the "Yutaka Takeda Hospital," a Municipality Hospital, and at the Parauapebas' Health Unit, Pará, from January to September 2006 and during March to November 2008. All samples were tested for rotavirus using immunochromatography, polyacrylamide gel electrophoresis (PAGE), and RT-PCR, yielding an overall positivity of 12.45% (30/241). Rotavirus G2P[4] was identified in 27 of 30 samples (90%), followed by G1P[8] (2/30, 6.67%) and G9P[8] (1/30, 3.33%). Phylogenetic analysis was performed in 15 of the G2 strains, all of which grouped into lineage II. Four of these strains clustered into sublineage II-a (year 2006) and 11 into one possible new sublineage named II-c (year 2008, except SAL-1920-C). The recent re-emergence of G2 genotype associated with lineage II in Brazil warrants the continuous monitoring of circulating rotavirus strains following the nationwide universal use of rotavirus vaccine.

Full genomic analysis and possible origin of a porcine G12 rotavirus strain RU172

Human group A rotavirus (GAR) G12 strains are regarded as potentially important pathogens for acute gastroenteritis. On the other hand, to date, the only report of detection of G12 in animals was that of a porcine G12P[7] strain RU172. Strain RU172 formed a separate G12 lineage, distinct from human G12 strains, and by analyses of deduced amino acid sequences, had a VP4, VP6, NSP4-5 of porcine origin. In the present study, we determined the full-length nucleotide sequences of VP1, VP3, and NSP1-3 genes and nearly full-length nucleotide sequence of VP2 gene of RU172. By nucleotide sequence identities and phylogenetic analyses, the VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5 genes of RU172 were assigned to G12-P[7]-I5-R1-C1-M1-A1-N1-T1-E1-H1 genotypes, respectively. Within their respective genotypes, (i) VP1 gene of RU172 exhibited higher genetic relatedness to Wa-like human G12 GARs than porcine strains, (ii) VP2-3 and NSP2 genes clustered separately from the Wa-like human (including G12) and porcine clusters, while (iii) the VP6, NSP1 and NSP3-5 genes clustered with porcine and porcine-like human strains. These observations suggested that (i) the porcine G12 strain might have originated from porcine-human reassortment events, or alternatively, (ii) the Wa-like human and porcine G12 strains might have originated from a common ancestor, and eventually evolved (by genetic drift and shift) with time. Our findings provided important insights into the possible patterns of evolution of the porcine G12 strain.

Identification of G8 rotavirus strains determined as G12 by rotavirus genotyping PCR: Updating the current genotyping methods

Background Rotaviruses are classified into G- and P-types, which are determined by the reactivity with antibodies to the outer viral proteins, VP7 and VP4, respectively, or sequence variation in the genes encoding these proteins. There are presently a number of different rotavirus strains co-circulating within the UK, with the common human strains G1P[8], G2P[4] and G9P[8] being the most prevalent. As part of strain surveillance for the European Rotavirus Network (EuroRotaNet) a cluster ( n = 29) of G8 strains was detected in the UK between February and May 2009. Objectives G8 strains were initially mistyped as G12 through multiplex RT-PCR, therefore further investigation was performed to ascertain the reasons behind this mistyping. Study design The genes encoding the VP7 of these G8 strains were sequenced and aligned with the existing G8- and G12-specific oligonucleotide primers. Results Multiple alignment of sequences derived from these strains and the G8- and G12-specific oligonucleotide primers revealed a series of point mutations which resulted in mismatches at the 3′ end of the G8-specific primer binding site that prevented amplification with the G8-specific primer, whilst a close homology with the G12-specific primer allowed mis-priming. Both the G8 and G12 primers were redesigned and their ability to correctly identify G8 and G12 strains was evaluated and confirmed. Conclusion These findings highlight the importance of monitoring the specificity and sensitivity of the genotyping methods in order to detect changes in the genotype distribution and changes associated with genetic drift of common or uncommon genotypes.

Phylogenetic analysis of human P[8]G9 rotavirus strains circulating in Brazil reveals the presence of a novel genetic variant

Background Group A rotavirus (RV-A) genotype P[8]G9 has emerged as one of the leading causes of gastroenteritis in children worldwide and currently is recognized as one of the five most common genotypes detected in humans. High intragenotype diversity in G9 RV-A has been observed, and nowadays, based on the genetic variability of the VP7 gene, six different phylogenetic lineages and eleven sublineages were described. Objectives To study the degree of genetic variation and evolution of Brazilian P[8]G9 RV-A strains. Study design Phylogenetic analysis of 19 P[8]G9 RV-A strains isolated from 2004 to 2007 in five different Brazilian states was conducted using the NSP1, NSP3, NSP5, VP4 and VP7 genes. For the VP4 and VP7 genes, 3D protein structure predictions were generated to analyze the spatial distribution of amino acid substitutions observed in Brazilian strains. Results Based on the phylogenetic analyses, all Brazilian strains clustered within lineage G9-III and P[8]-3 for VP7 and VP4, respectively, and were classified as genotype A1, T1 and H1 for the NSP1, NSP3 and NSP5 genes, respectively. Interestingly, all the strains isolated in Acre State (Northern Brazil) formed a closely related cluster clearly separated from the other Brazilian and prototype strains with regard to the five genes studied. Unique amino acid substitutions were observed in Acre strains in comparison with the prototype and Brazilian strains. Conclusion Inclusion of Acre strains in the phylogenetic analysis revealed the presence of a novel genetic variant and demonstrated a diversification of P[8]G9 rotaviruses in Brazil.

Identification of a Rotavirus G12 Strain, Indonesia

To the Editor: Group A rotaviruses are the most common etiologic agents of acute gastroenteritis in infants and young children, each year resulting in ≈100 million diarrhea episodes and 600,000 deaths worldwide (1). The genome of rotavirus comprises 11 segments of double-stranded RNA, which encode 6 structural viral proteins (VPs) and 6 nonstructural proteins (NSPs). Recent scientific reports have identified novel rotavirus strains, such as G12 (2–5), which were first described in 1987 among Filipino children with diarrhea (6). In Indonesia, a rotavirus study showed that a broad variety of VP7 types (G1, G2, G3, G4, G8, G9) and VP4 types (P[4], P[6], P[8], P[9], P[10], P[11]), especially G9 and P[8] and G9P[8], were the genotype combinations most frequently encountered (7). From 2005 through 2008, we conducted a nationwide surveillance study among children who had diarrhea to determine etiologies among Indonesian children seeking health services for diarrhea at hospitals and health clinics. Patients were enrolled after obtaining consent from parents/guardians of those eligible in accordance with an institutional review board protocol approved by the US Naval Medical Research Unit No. 2 (NAMRU-2) and the Ethical Committee of the Indonesian National Health Research and Development Institute. Stool specimens and clinical enrollment data were collected for each eligible patient, and all collected items were transported to NAMRU-2 in Jakarta, Indonesia. In December 2007, a stool specimen was collected from a 14-day-old afebrile infant brought to Sumber Waras Hospital in West Jakarta with diarrhea, vomiting, moderate dehydration, and malnutrition. This patient was infected with the rotavirus G12 strain, was hospitalized for 6 days, and was discharged after recovering fully. Bacterial cultures and ova/parasite evaluations were negative for enteric pathogens. Rotavirus was detected in this specimen and genotyped by multiplex, seminested reverse transcription–PCR (RT-PCR) targeting the VP4 and VP7 genes (8,9). The specimen was typed as P[4]P[6] but was G-nontypeable. Primers to detect G12 were then used for RT-PCR and identified the proper G12 amplicon size (2,3). By use of published primers (9), sequencing of the VP7 gene segment confirmed the presumptive G12 genotype. Sequencing reactions were performed by using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA) on the Applied Biosystems 3130xl sequencer. Sequence analysis was done by using Sequencher 4.8 version (Gene Codes Corporation, Inc., Ann Arbor, MI, USA). Nucleotide sequences were submitted to GenBank for a BLASTN search (http://blast.ncbi.nlm.nih.gov) on the National Center for Biological Information website. We then created alignments of nucleotides and deduced amino acid sequences and compared them with a selection of reference strains from the GenBank database. Genetic relationships among G12 were determined by using PAUP version 4.0 beta 10 software (http://paup.csit.fsu.edu). A phylogenetic tree was constructed on the basis of nucleotides 1–971 of the VP7 gene by using the neighbor-joining method and applying the Kimura 2-parameter method with 1,000 bootstrap replicates of the neighbor-joining model. The BLASTN search of the VP7 989 nucleotide sequence of the putative G12 Indonesian rotavirus (Indo SWJ0806) showed 98% similarity with published VP7 sequences of rotavirus G12 strains from Japan (CP727; GenBank accession no.{type:entrez-nucleotide,attrs:{text:AB125852,term_id:49065032,term_text:AB125852}}AB125852), Argentina (Arg721; GenBank accession no. {type:entrez-nucleotide,attrs:{text:EU496255,term_id:169835103,term_text:EU496255}}EU496255), and Thailand (T152; GenBank accession no. {type:entrez-nucleotide,attrs:{text:AB071404,term_id:19912798,term_text:AB071404}}AB071404). The Indonesia G12 clustered into the lineage II composed of rotavirus G12 reference strains from Japan, Argentina, South Korea, and Thailand (Figure). Lineage II is a minority cluster when compared with lineage III, which consists of rotavirus G12 from the United States (US6588, Se585), Saudi Arabia (MD844), India (13B2), Bangladesh (RV161), and other Thailand strains (MS051) (4). The nucleotide sequence divergence between lineage II and lineage III ranges from 2.6% to 3.2%. Analysis of the deduced amino acid sequence alignment on the neutralization epitopes that code for the antigenic regions A, B, and C show high conservation of the most immunodominant sites (data not shown). Antigenic regions A, B, C, D, E, and F of Indonesia SWJ0806 show 100% amino acid similarity to Japan G12 strains; K12 and CP727 (9). The amino acid residue at position 142 of the antigenic region B has characterized lineage I and II (Val) and lineage III (Leu). Figure Phylogenetic analysis of the viral protein (VP) 7 genotype G12 rotavirus of Indonesia with reference strains downloaded from GenBank. The GenBank accession numbers of each strain appear next to the strain. The multiple alignment was constructed by using ... Phylogenetic analysis showed that the virus clusters into lineage II and that the deduced amino acid sequence is highly conserved compared with other reported rotavirus G12 strains identified. The combination of the P[6] genotype in this rotavirus strain suggests the possibility of a zoonotic transmission (10). Continued surveillance for rotavirus is an essential component of a country’s public health infrastructure and diarrhea prevention programs. Rotavirus genotyping from the data obtained provides necessary information for vaccine development and identification of novel and emerging rotavirus strains.