Recombinase polymerase amplification technology for point-of-care diagnosis of neglected tropical diseases
Luisa Hueso, Sara Martorell, Amadeo Sena‐Torralba, Maite Ferrando, Mireia Ferri, Ángel Maquieira, Francine Ntoumi, Sergi Morais
Abstract
• RPA enables rapid, isothermal diagnosis of neglected tropical diseases (NTDs). • Ideal for point-of-care testing in decentralized, low-resource settings. • Freeze-dried RPA reagents eliminate cold-chain needs for improved accessibility. • RPA faces primer design challenges but suits field diagnostics globally. • RPA integration with portable platforms supports field-based NTD diagnostics. Recombinase Polymerase Amplification (RPA) technology significantly advances the diagnostics of neglected tropical diseases (NTDs), providing rapid, isothermal, and minimally preparative testing ideally suited for under-resourced countries. This review critically assesses the current applications, limitations, and potential of RPA for detecting a broad spectrum of NTD pathogens, including viruses, bacteria, helminths, and fungi. The ability of RPA to operate under constant temperature conditions without the need for complex thermal cycling facilitates rapid pathogen detection within minutes, enhancing its utility for decentralized point-of-care testing in remote and underserved regions. RPA, however, faces limitations, including the labor-intensive and costly validation of primer design, especially for multiplex assays, and a susceptibility to nonspecific amplification. These challenges highlight the need for continuous refinement to ensure reliable and consistent performance across diverse environmental conditions. Despite these constraints, the scalability of RPA assays and their compatibility with portable detection platforms make them well-suited for deployment in field settings without access to traditional laboratory infrastructure. This review emphasizes the transformative potential of RPA in NTD diagnostics, enhancing accessibility, precision, and timeliness of interventions, ultimately contributing to improved global public health outcomes.