Abstract
Group A rotaviruses (RVAs) are the most important etiological agent of acute gastroenteritis in children <5 years of age worldwide. The monovalent rotavirus vaccine Rotarix was introduced into the national Expanded Programme on Immunization (EPI) in Ghana in May 2012. However, there is a paucity of genetic and phylogenetic data on the complete genomes of human RVAs in circulation pre-vaccine introduction. The common bovine rotavirus VP7 genotype G8 has been sporadically detected in Ghanaian children, usually in combination with the VP4 genotype P[6]. To investigate the genomic constellations and phylogeny of RVA strains in circulation prior to vaccine introduction, the full genomes of two unusual G8P[6] strains, GH018-08 and GH019-08, detected during burden of disease surveillance, were characterized by Illumina MiSeq sequencing. The Ghanaian isolates, GH018-08 and GH019-08, exhibited the unusual, previously unreported genotype constellation G8-P[6]-I2-R2-C2-M2-A2-N2-T2-E2-H3. Phylogenetic analyses confirmed that 10 out of the 11 genes of GH018-08 and GH019-08 were identical/nearly identical, with significant variation detected only in their VP1 genes, and clearly established the occurrence of multiple independent interspecies transmission and reassortment events between co-circulating bovine/ovine/caprine rotaviruses and human DS-1-like RVA strains. These findings highlight the contribution of reassortment and interspecies transmission events to the high rotavirus diversity in this region of Africa, and justify the need for simultaneous monitoring of animal and human rotavirus strains.
Highlights
Group A rotaviruses (RVAs) are the most important etiological agent of acute gastroenteritis in children,5 years of age worldwide, and are estimated to be responsible for most of the 453,000 rotavirus-associated deaths annually, of which the majority occur in countries of sub-Saharan Africa and South East Asia [1]
GH018-08 and GH019-08 had been screened for RVA by Enzyme Immunoassay (EIA) using the commercial IDEIA enzyme immunoassay kit (Dako Diagnostics Ltd., Cambridgeshire, UK), and polyacrylamide gel electrophoresis (PAGE) of RNA as previously described [38]
Nucleotide sequencing and assignment of genotypes Full-length or nearly full-length sequences of all 11 genomic segments of GH018-08 and GH019-08 were successfully determined by Illumina MiSeq sequencing
Summary
Group A rotaviruses (RVAs) are the most important etiological agent of acute gastroenteritis in children ,5 years of age worldwide, and are estimated to be responsible for most of the 453,000 rotavirus-associated deaths annually, of which the majority occur in countries of sub-Saharan Africa and South East Asia [1]. The traditional binary classification system for RVAs is based on the characteristics of the two outer capsid proteins VP7 (glycoprotein, G-genotype), and VP4 (proteasesensitive protein, P-genotype) [3], [4]. This classification system cannot comprehensively define molecular epidemiology and evolutionary relationships among circulating RVA strains. An extended RVA classification and nomenclature system has been established based on the assignment of genotypes to all 11 genome segments [3], [5]. While at least 73 G/P genotype combinations have been described in humans [7], only a limited number of strains - G1P[8], G2P[4], G3P[8], G4P[8], G9P[8], and to a lesser extent G12P[8] and G12P[6] - are most frequently associated with the RVA disease burden globally [8,9,10,11]
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