Camille Escadafal, Pierre Garneret, Jean-Claude Manuguerra, Anavaj Sakuntabhai, Patrick Tabeling, Pierre Lafaye, Aurélia Kwasiborski, Jessica Vanhomwegen, Fabrice Monti, Beatrice Jacquelin, Laura Magro, Gulliver (UMR 7083), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), HIV, Inflammation et persistance, Institut Pasteur [Paris], Cellule d'Intervention Biologique d'Urgence - Laboratory for Urgent Response to Biological Threats (CIBU), Génétique fonctionnelle des Maladies infectieuses - Functional Genetics of Infectious Diseases, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Ingénierie des Anticorps (plate-forme) - Antibody Engineering (Platform), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], The authors acknowledge financial support from the Pasteur Ebola Task Force directed by Pierre Legrain, and from Carnot Pasteur Maladies Infectieuses (ANR 11-CARN 017-01)., Sincere thanks also go to all the institutions that made the trip to Guinea possible: Pasteur Institute International Network, French and Guinean Red Crosses. For the access to the Macenta laboratory and all the administrative agreements, the authors thank Sylvain Baize, Kathleen Victoir, Chloé Pelicier, Mialy Rabenoro, Virginie Pirard, Cassandre von Platen and Nathalie Jolly. The authors are grateful to the patients, clinicians and the staff from the Ebola Treatment Center for their participation and efficient collaboration. The authors thank MicroFactory and R&DVision for the fabrication of transportable detection equipment, in a very short time., HIV, Inflammation et persistance - HIV, Inflammation and Persistence, Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
The most performing techniques enabling early diagnosis of infectious diseases rely on nucleic acid detection. Today, because of their high technicality and cost, nucleic acid amplification tests (NAAT) are of benefit only to a small fraction of developing countries population. By reducing costs, simplifying procedures and enabling multiplexing, paper microfluidics has the potential to considerably facilitate their accessibility. However, most of the studies performed in this area have not quit the lab. This letter brings NAAT on paper closer to the field, by using clinical samples and operating in a resource-limited setting. We first performed isothermal reverse transcription and Recombinase Polymerase Amplification (RT-RPA) of synthetic Ribonucleic Acid (RNA) of Ebola virus using paper microfluidics devices. We further applied this method in Guinea to detect the presence of Ebola virus in human sample RNA extracts, with minimal facilities (carry-on detection device and freeze-dried reagents on paper). RT-RPA results were available in few minutes and demonstrate a sensitivity of 90.0% compared to the gold-standard RT-PCR on a set of 43 patient samples. Furthermore, the realization of a nine-spot multilayered device achieving the parallel detection of three distinct RNA sequences opens a route toward the detection of multiple viral strains or pathogens.