Prevalence and Genetic Diversity of Entamoeba histolytica Strains Isolated from Patients in Health Facilities in the Buea Health District

Abstract

Aims: Amoebiasis remains a significant cause of diarrheal disease in regions with limited sanitation. In this study, we aimed at investigating the prevalence and genetic diversity of Entamoeba histolytica strains circulating in the Buea Health District. Study Design: A cross-sectional study. Place and Duration of Study: This study was carried out in 8 health facilities in the Buea Health District (BHD), Cameroon, between January 2024 and June 2024. Methodology: Overall, 265 participants (162 females and 103 males) aged 1–90 years were randomly selected from eight health facilities in the Buea health district. Stool samples were collected and examined for the presence of Entamoeba histolytica (E. histolytica) cysts using the formol-ether concentration technique and wet mount microscopy. DNA was extracted and polymerase chain reaction targeting the SSU rRNA gene was done for all microscopy-positive samples for confirmation. Genetic diversity was investigated using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Data analysis was performed using R software version 4.2.3 (RCore Team, 2023) and statistical significance was set at P < .05. Results: The prevalence of Entamoeba histolytica in the BHD was 11.7%. Of the 265 samples, 35 were positive by microscopy and 31 microscopy positive samples were confirmed positive by PCR. The prevalence was significantly higher among children aged 1–5 years (27.04%, P = .02). Female participants recorded a slightly higher prevalence of 12.35% compared to males (10.68%), although this difference was not significant (P = .08). The highest prevalence was obtained from samples collected from the Tole Integrated Health Center. The serine-rich E. histolytica protein was amplified in 28 samples, Out of these, 5 different band profiles were obtained following AluI digestion of the nested-PCR serine-rich E. histolytica protein products. Conclusion: These findings confirmed notable prevalence and the genetic heterogeneity of E. histolytica in the district. This diversity could influence clinical presentation and complicate control efforts, underscoring the need for strain-specific surveillance using more advanced genotyping methods, as PCR-RFLP provides limited information about genetic variation. Further work is needed using more robust molecular techniques.