FILODIAG

There is an urgent need for fast and accurate diagnostic tests in the current Ebola crisis. The rapid diagnosis of EVD during early and late stage of infection is a decisive step for risk assessment and for guidance to physicians to take the necessary decisions to limit the spread of the infection, and to safely nurse the infected patients. While fast and easy-to-use tests usually rely on immuno-diagnostic approaches, they typically lack high sensitivity and specificity. The gold standard for accurate diagnostics, (Real-Time) PCR, however, is limited in terms of processing time, high costs, and in most cases also in terms of ease of use and the need of special laboratory facilities. The aim of this project is to provide an ultra-fast and accurate molecular diagnostic instrument that can perform a molecular Ebola test within 10-15 minutes. Patients, suspected of Ebola Virus Disease (EBD), due to fever and/or other clinical signs, could be rapidly and accurately tested at the Point-of-Care by withdrawal of blood, or less invasive fluids, such as saliva or urine. The core technology of this proposal is based on laser-heated nanoparticles (Laser PCR), overcoming the time-limiting step of heating and cooling the reaction sample in conventional PCR allowing for drastically reduced turn-around times. The project partners (GNA, INMI, MUB, EMG) have the competence to integrate sample preparation with virus-binding magnetic particles, ultra-fast molecular diagnostics with Laser PCR, test the system in a biosafety level 4 laboratory in Rome, Italy, and thoroughly conduct field testing in a medical care centre in West Africa (Sierra Leone). A tentative Ebola Laser PCR assay has already been set up, detecting 10 target copies of synthetic DNA oligos, corresponding to the Ebola genome sequence, in less than 12 minutes. The FILODIAG project will deliver a potentially multiplexed diagnostic system fast enough for point-of need testing of incoming patients at low-resource