Molecular Characterization of Rotaviruses From A SemiUrban Communication in The Central Region of Ghana

ABSTRACT

Acute gastroenteritis (AGE) is an inflammation of the intestinal mucosa and is characterised by the onset of diarrhoea with or without vomiting, fever, and abdominal pain. Rotaviruses are the leading cause of AGE in infants and young children resulting in severe dehydration, which can eventually lead to hospitalisation and death. In Ghana, diarrhoea is implicated in the death of about 11% of all children under the age of 5 years, with rotavirus being the most responsible causative agent as with the case of most countries in the world. Currently, in Ghana, the prevalence of rotavirus is 28%. Rotavirus vaccine, Rotarix has been introduced into the immunisation system in Ghana to reduce morbidity and mortality due to the virus. This study was therefore designed to determine the prevalence of rotaviruses in a semi-urban setting, post vaccine introduction era. Stool samples collected from children under five years of age with AGE, presenting at various Health Centres in the Breman Asikuma area in the Central Region of Ghana were used. The samples were tested for Group A rotaviruses using the ProwflowTM rotavirus enzyme Immunoassay (EIA) kit. Viral RNA was extracted from all samples using the Phenolchloroform extraction method and electrophoresed on polyacrylamide gel for group characterization as well as the determination of RNA integrity. Negative EIA stool samples were then subjected to NSP3 qRT-PCR. Rotavirus genotypes of positive samples were determined by semi-nested RT-PCR with both gene and genotype specific primers, and the genotypes were further confirmed by Sanger sequencing. Further characterization of the virus was done by whole genome sequencing on the Illumina platform. Results showed that out of thirty-six (36) samples collected, three (9.0%; 3/36) tested positive by EIA. Electrophoretic analysis of the samples, also showed that two (2) of the samples were of the long electrophoretype and one (1) sample is of the short electrophoretype. Circulating strains were found to be G9P[8] and G3P[6]. Full genome characterization of the selected strains gave Walike constellations. The results of the study suggest that AGE due to rotavirus is relatively lower compared to the national incidence reported in literature.