Rotavirus is associated with severe infantile diarrhoea requiring hospitalization. In order to
control severe disease caused by rotavirus infection, vaccination is considered an essential
strategy. Currently, two rotavirus vaccines (Rotarix™ and RotaTeq™) have been included in the
Expanded Program on Immunization of most African countries to help reduce the high disease
indections. Ghana has over the past 20 years been involved in the surveillance of rotavirus
diseases by isolating rotaviruses from diarrhoeic stools and determining the strain types as part
of the build up to vaccine introduction. During rotavirus burden of disease surveillance between
2007 and 2011, three hundred and one rotavirus samples could not genotype with the pool of
primers recommended for use within the WHO Regional Reference Laboratory network.
Samples with sufficient stools were characterized.
The study sought to characterize and identify primer mismatches for nontypeable rotavirus
strains at the Regional Rotavirus Reference laboratory in Ghana.
One hundred and eleven G and 89 P-types had sufficient stool and were characterized by
sequencing methods. To confirm previous EIA results, 10% of randomly selected nontypeable
samples were reanalyzed using the commercially available DAKO IDEIA kit (Dako Diagnostics,
Cambridgeshire, UK). To assess the integrity of RV dsRNA, Polyacrylamide Gel
Electrophoresis (PAGE) was carried out. Viral ribonucleic acids were extracted by the phenolchloroform-guanidine-isothiocyanate method and one step RT-PCR targeting the VP7 and VP4
genes of RVA was carried out using PCR primer pairs, Beg9/End9 and 9Con1/9Con2 for VP7 genes and Con2/Con3 and VP4F/VP4R for VP4 genes. The RT-PCR products were purified with
the QIAquick PCR purification kit (Qiagen/Westburg), sequenced with the ABI PRISM®
BigDye Terminator v.3.1 Ready Reaction mix. Amplicons were purified using ethanol-sodium
acetate precipitation method. Sequences were subjected to BLAST (Basic Local Alignment
Search Tool) and subsequently characterized using the web-based genotyping tool, RotaC v2.0.
The G-types were determined for 74 out of the 111 samples that had sufficient stool to work
with. The G-types detected were as follows: G1; 31 (41.8%), G2; 9 (12.1%), G3; 11 (14.8%),
G6; 1 (1.4%), G8; 2 (2.7%), G9; 14 (18.9%) and G12; 6 (8.1%). Sequence analyses of the
Ghanaian VP7 isolates and cognate genes of known ancestral and contemporary reference strains
revealed the accumulation of point mutations in the antigenic regions.
The P-types were successfully characterized for 57 of the 89 samples that had sufficient stool.
Forty-eight (84.2%) of these were identified as Pa strains of which 5 (10.4%) were
characterized as the rare OP354-like human rotavirus Pb subtype. Other strains detected in the
study were P, 1 (1.8%), P, 7 (12.3%) and P, 1 (1.8%). P was found in combination
with G6. In Silico PCR performed with in-house primers failed to genotype Ghanaian P
isolates. However, In Silico PCR with published primers, Rev compl 1T-1D and P8b-MMC38
successfully genotyped the Ghanaian Pa and Pb isolates respectively
CDR, C (2021). MOLECULAR CHARACTERIZATION OF PREVIOUSLY NON-TYPEABLE ROTAVIRUS STRAINS IN GHANA. Afribary.com: Retrieved April 15, 2021, from https://afribary.com/works/molecular-characterization-of-previously-non-typeable-rotavirus-strains-in-ghana
Coalition, CDR. "MOLECULAR CHARACTERIZATION OF PREVIOUSLY NON-TYPEABLE ROTAVIRUS STRAINS IN GHANA" Afribary.com. Afribary.com, 02 Apr. 2021, https://afribary.com/works/molecular-characterization-of-previously-non-typeable-rotavirus-strains-in-ghana . Accessed 15 Apr. 2021.
Coalition, CDR. "MOLECULAR CHARACTERIZATION OF PREVIOUSLY NON-TYPEABLE ROTAVIRUS STRAINS IN GHANA". Afribary.com, Afribary.com, 02 Apr. 2021. Web. 15 Apr. 2021. < https://afribary.com/works/molecular-characterization-of-previously-non-typeable-rotavirus-strains-in-ghana >.
Coalition, CDR. "MOLECULAR CHARACTERIZATION OF PREVIOUSLY NON-TYPEABLE ROTAVIRUS STRAINS IN GHANA" Afribary.com (2021). Accessed April 15, 2021. https://afribary.com/works/molecular-characterization-of-previously-non-typeable-rotavirus-strains-in-ghana