The decentralization of laboratory capacities is a vital strategy for enhancing epidemic preparedness and response across Africa. Centralized laboratory systems often lead to delays in case confirmation, which can hinder timely interventions and worsen the impact of outbreaks, particularly in rural and hard-to-reach areas. The newly released guidelines provide a structured approach to decentralization, emphasizing strategic goals such as strengthening laboratory capacities at subnational levels. This ensures timely detection and control of diseases prone to epidemics. Key guiding principles include equity, country ownership, multisectoral collaboration, and evidence-based decision-making. The implementation framework outlines practical steps for planning, executing, and sustaining decentralized diagnostic networks, along with mechanisms for intra- and post-implementation reviews to facilitate continuous improvement. Furthermore, these guidelines advocate for integration with existing surveillance systems, case management, and infection prevention and control (IPC) measures, focusing on the One Health approach. While informed by the Mpox outbreak response, the guidelines are adaptable to other priority diseases and align with the International Health Regulations (IHR 2005) and the Africa CDC Strategic Plan (2022-2027).