<p>Nigeria has recorded recurrent Mpox outbreaks since the 1970s, with the most recent resurgence underscoring weaknesses in the country’s diagnostic and laboratory infrastructure. Despite improvements in molecular diagnostics, such as genome sequencing and polymerase chain reaction (PCR), limited access to testing facilities, shortages of trained personnel, and inadequate biosafety measures remains a significant challenge. Although collaborations with international partners during recent outbreaks of infectious diseases have enhanced diagnostic capacity. The COVID-19 pandemic led to investments that expanded diagnostic infrastructure and decentralized sample collection, yet logistical barriers persist, particularly in rural areas where inadequate transportation networks and power supply disruptions delay sample processing. Additionally, workforce shortages and high emigration rates among laboratory personnel further weakened the system’s efficiency. Therefore, this review evaluates Nigeria’s preparedness for Mpox surveillance and response by analyzing past outbreak management efforts, including Ebola and Lassa fever. It identifies key strengths and gaps in the laboratory system, highlights barriers to effective diagnostics, and explores opportunities for improvement in terms of upgrading biosafety infrastructure, capacity building of healthcare workers, fostering public-private partnerships to develop local diagnostic tools, and increasing government funding for laboratory services. Strengthening these areas is essential for improving Nigeria’s capacity to detect and contain Mpox outbreaks and enhancing overall public health preparedness.</p>