Development of a recombinase polymerase amplification-based point-of-care detection system for mpox

Mpox virus (MPXV) is a zoonotic virus from the genus Orthopoxvirus of the Poxviridae family and is the causative agent of Mpox disease. Since the beginning of 2025, the disease was responsible for a total of 24,672 confirmed cases and 82 fatalities and is present in 75 countries. MPXV is regarded as a virus with major pandemic risk and has been designated by the World Health Organization (WHO) as a Health Emergency of International Concern (PHEIC). Effective local monitoring and surveillance of epidemic sites require portable diagnostic tools capable of rapid and accurate detection of the pathogen. The tendency of co-infection, as well as the high variance in symptoms between individuals, makes the disease hard to diagnose by basic superficial examinations. For this reason, molecular diagnostic tools present a popular and accurate alternative to traditional diagnostic methods. PCR stands out as the gold standard in molecular detection. However, the components and the environmental conditions needed to conduct a PCR test necessitate the presence of specialized lab equipment and a sterile working environment. RPA is an alternative molecular amplification and detection tool that is popular within point-of-care (PoC) applications. Similar to PCR, RPA uses only two primers, works with similar samples, and has comparable sensitivity and specificity. In line with this, we present an RPA-based molecular detection system, capable of detecting target genetic material at 2.92167 x 104 copies. The system was integrated into a portable, PoC device capable of running the reaction and providing all necessary components for signal detection using a smartphone camera. Together, the platform provides a rapid, portable, and accessible diagnostic solution for Mpox, supporting improved outbreak surveillance and timely clinical decision-making in resource-limited settings.