Abstract
BACKGROUND: Diarrhea is a leading cause of mortality in children under five, with rotavirus and enteric adenoviruses F40/F41 being significant contributors. In regions like Africa and Southeast Asia, the incidence is exacerbated by poor hygiene and sanitation, as well as the challenges around availability and affordability of healthcare and disease diagnostics. The objective of this systematic review was to identify and evaluate molecular assays for the detection of rotavirus and enteric adenoviruses F40/F41, with particular emphasis on diagnostic performance and suitability for point-of-care use. METHODS: A systematic review of literature from 2001 to 2024 was conducted following PRISMA guidelines. Out of 659,613 records screened, a subset of primary studies reporting on molecular assays development or diagnostic performance met the inclusion criteria. Studies exclusively focused on epidemiology, vaccine impact, or narrative reviews were excluded from the analytical synthesis. RESULTS: The review identified 26 PCR-based assays and 3 isothermal amplification assays. Reported clinical sensitivities ranged from approximately 85% to 100%, with specificities generally above 95% across studies. Several assays also demonstrated low limits of detection and turnaround times compatible with decentralized use. In particular, three LAMP assays and three PCR-based methods fulfilled key performance and operational criteria for point-of-care diagnostics. The review concluded that molecular diagnostics, especially LAMP assays, are effective for rapid detection of rotavirus and enteric adenoviruses F40/F41. CONCLUSION: While PCR assays are considered the gold standard, LAMP assays are more accessible and faster, making them better suited for use in resource-limited settings. These assays show promising performance and may support point-of-care diagnostics. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-026-13081-4.