Periodic Reporting for period 3 - EbolaMoDRAD (Ebola Virus: Modern Approaches for developing bedside Rapid Diagnostics – Sofia ref.: 115843)
Reporting period: 2017-01-01 to 2018-01-31
-to develop a biosafe detection method (diagnostic tool) for use at points of care in EVD endemic countries;
-to validate the diagnostic tools first in BSL4 and reference laboratories and then in the field;
-to implement a strong capacity building programme in West Africa with focus on rapid diagnostic, biosafety measure and outbreak management;
-to disseminate widely the project and its results to public health bodies, NGO, soutbreak management teams and local hospitals in West Africa.
Within this period, successful progress was made to achieve this objective. By using Ebola blood samples (from NHP), we could confirm our earlier data on our recently developed inactivation vacuum tube for Ebola virus. We also succeeded in pushing our isothermal assays towards two final versions that could be used in the diagnostic laboratory or field setting in future outbreaks. We have identified conventional real-time PCR assays that are most suitable as Ebola diagnostics and we are now looking to make these into formats that can be deployed in the field. We developed over 100 cartridges which contain the reagents and a silicon chip in order to diagnose Ebola infection. In addition, we developed and validated immunochromatographic flow tests (LFD) with a sensitivity of 89% and specificity of 98% by using clinical samples in Africa and Europe. We are now exploring possibilities to bring this assay to the marke and which can be used in future outbreaks.
EbolaMoDRAD has investigated the influence of the extraction reagents from the direct inactivation blood tubes on the real-time PCR and also development of these reagents into freeze-dried reagents that can be easily stored for extended periods of time. Regarding the isothermal diagnostic development, the direct novel RNA detection approach has a strong potential to be patented for use in molecular diagnostics. This will serve as a method for RNA-based detection, opening new doors for diagnostic approaches. We also developed strips capable of detecting the RPA-based Ebola diagnostic. These strips offer the potential of instrument-free molecular amplification with a simple lateral flow read-out. This type of diagnostic could sit well alongside established conventional serological diagnostic test strips. The development of the Clonit complete sample to answer system has significantly progressed. Extraction of clinical samples for molecular testing still remains an Achilles heel for many molecular diagnostics as these systems are both column and centrifuge-based, or rely on very expensive robotic machinery. AJ Innuscreen has made great advances in the field of extraction, particularly with a bespoke solution of automated nucleic acid extraction. We have shown that the production of recombinant EBOV proteins enables the development of highly specific serological assays for analysing immune responses against EBOV. The assay can be developed for diagnosing acute infections and analysing EBOV vaccine-induced responses as well as analysing the seroprevalence against EBOV in larger population studies. LFA antigen and IgM detection has been developed and partly validated, these tools together will contribute to a better rapid field diagnostic. All developed diagnostic tools for a rapid diagnosis of Ebola infection developed were validated, representing one of the successes of the project. Collaborations between several partners beyond the objective of this project were established, which may lead to new initiatives for development of diagnostic tools for neglected pathogens.
EbolaMoDRAD publications: http://www.ebolamodrad.eu/index.php/press-and-publications
Project coordinator: Ali Mirazimi, FoHM
Contact details: Ali.Mirazimi@folkhalsomyndigheten.se