Molecular Detection and Sequencing of Rotavirus VP4 among Children Aged 0-5 Years with Gastroenteritis in 2 Selected Healthcare Centres in Keffi, Nigeria

Aims: This study was conducted to detect and sequence Rotavirus VP4 among children aged 0-5 years with gastroenteritis in 2 selected healthcare centres in Keffi, Nigeria.

Study Design: The study was a cross sectional study.

Place and Duration of Study: Keffi, Nasarawa State, between March and June 2019.

Methodology: Stool samples were collected from 303 (203 from FMC Keffi and 100 from PHC Angwan Waje, Keffi) children with gastroenteritis and information about them were obtained by structured questionnaires. All collected samples were screened for the presence of Rotavirus antigen using Aria Rotavirus antigen detection test kit (CTK Biotech, Inc, San Diego, USA). VP4 was detected from Rotavirus positive samples by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) using specific primers. The sequences of the amplified VP4 genes were verified using MEGA software version 7 and Rotavirus strains were determined by pasting the FASTA (Text based format for representing nucleotide sequence) format into the Basic Local alignment Search Tool (BLAST) from the National Center for Biotechnology Information (NCBI). Data collected were analysed using Smith’s Statistical Package (version 2.8, California, USA) and P value of ≤ 0.05 was considered statistically significant.

Results: Of the 303 children screened, 54(17.8%) were positive for Rotavirus infection. Highest prevalence of the viral infection was recorded among males (19.7%) aged 0-12 months (24.8%). Gender was found to be associated with rate of Rotavirus infection in this study (P<0.05). However, age was not significantly associated with the viral infection (P>0.05). Furthermore, based on the RT-PCR carried out, 3(5.6%) out of the 54 Rotavirus positive samples were positive for the VP4 gene and sequences of this gene were all found to be of type P [11] Strain N115.

Conclusion: This study reveals the presence of infection with type P [11] Strain N115 (5.6%) of Rotavirus in the study population. The detection of this rare rotavirus strain in this study is a cause for concern and hence there is an urgent need for the Nigerian health authorities to implement a nationwide surveillance system for monitoring rotavirus molecular epidemiology.