Community Research and Development Information Service - CORDIS

H2020

EbolaMoDRAD Report Summary

Project ID: 115843
Funded under: H2020-EU.3.1.7.13.

Periodic Reporting for period 2 - EbolaMoDRAD (Ebola Virus: Modern Approaches for developing bedside Rapid Diagnostics)

Reporting period: 2016-01-01 to 2016-12-31

Summary of the context and overall objectives of the project

Within the second period of the project we have developed several assays and protocols, which will facilitate achievement of EbolaMODRAD overall goals. We have, among others, successfully developed an inactivation vacuum tubes, which not only contribute to a safer transfer and shipment of samples, but also allows the samples to be handled outside of the high containment laboratory, which means easier and more rapid diagnoses for patients. These data has been published (Rosenstierne MW et. al. 2016) and has been positively received by the infectious disease community. We have also succeeded to develop several molecular diagnostic tools which has been validated or in progress for validation with human samples.
Within the second period of the project, we have also successfully developed several tools such peptides, antibodies and recombinant proteins, which has been and will can be used for development of serological diagnostic tools. We have also succeeded to develop an immunochromatographic lateral flow tests (LFD) which has been evaluated by in field assays in Sierra Leone.
We have managed to establish a project Biobank for linked with epidemiological and clinical data, to be used for research purposes. These samples has been used for validation of new assays at the field.
Within the second period of project we have even initiated and performed a workshop focusing on building a cooperation. An exchange platform on mobile laboratory deployment for outbreak investigations has also been organised.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The project is organised into five work packages (WP): the first one is dedicated to the management of the consortium and the organisation of annual and Executive Committee (ExCom) meetings.
The WP2 team has achieved great success this year. Our work on inactivating Ebola directly in blood tubes has been published (Rosenstierne MW et. al. 2016) and has been positively received by the infectious disease community. Considerable progress has been made in development of a single linear RCA assay, following the decision in January 2016 to move over from the hRCA approach due to problems with non-specific amplification and production of double-stranded snippets. The RPA assays have undergone significant development. Both Coris and the University of Stirling have made impressive progress, making a transition from the original fluorescence based assays into a triplex lateral flow molecular diagnostic. Two independent partners (AMU and UH) have tested and identified conventional real-time PCR assays that are most suitable as Ebola diagnostics and are now looking to make these into formats that can be deployed in the field. Clonit have developed over 100 cartridges which contain the reagents and a silicon chip in order to diagnose Ebola infection.
In WP3, the summary can be divided within the tasks.
All EBOV genes as well as GP genes were successfully expressed by baculovirus expression system with and without His-tag. Purified proteins have been used to immunize rabbits. Further work is ongoing for their further characterization, expression of MARV, SUDV, and BDBV protein.
Scan peptides covering all Ebolavirus proteins targeted of Zaire ebolavirus have been successfully synthesized. After further optimization of the print conditions, the microarray was used for the characterisation and epitope mapping of several commercial anti-VP40 antibodies from BBI solutions that had been employed in a prototype LFD assay by CORIS as well as for a complete epitope mapping of anti-EBOV pAbs produced by UTU.
First immunochromatographic flow tests (LFD) have been developed by CORIS with antibodies from commercial sources. 5 of these pre-prototypes have been evaluated with live viruses in BSL-4 laboratories and 100 tests of these two LFD antigen detection test prototypes have been evaluated in field assays in Sierra Leone. Based on the evaluation results, a 2nd generation lateral-flow antigen detection prototype called “Boosted version” was developed. It showed a sensitivity of 89% and specificity of 98% in further field trials with a total number of 250 tests.
The general objectives of WP4 are to validate analytic sensitivity and specificity of new diagnostic tools both in BSL4 labs in Europe and in the field using samples included in a “disseminated” BioBank created in the framework of this project. Specifically, efficiency and operational evaluation and proficiency test of the new diagnostic tools will be performed on panels of Ebola virus positive and negative samples assembled from the repository.
Within the WP5, a workshop focusing on building a cooperation and exchange platform on mobile laboratory deployment for outbreak investigations has been organised. A third one is planned, aiming to disseminate knowledge on outbreak management based on the experience from latest Ebola virus outbreak.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

In WP2, we are now investigating 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 which make them much more applicable for tools in outbreaks. This progress is bringing together sample collection right through to patient results in a safe and rapid manner.

Partners in WP3 has expressed and purified EBOV VP24, VP30, VP35, VP40 and NP genes as GST-fusion proteins and conducted complete synthesis of corresponding scan peptides and fabricated a novel microarray screening platform. This platform are being used for (1) full characterization of produced polyclonal antibodies for LFD assay developments, and (2) screening of human and primate sera.
Regarding WP4, the setting up of a “centralized” BioBank should have had the purpose to create a platform to facilitate future research activities. To date, due to the difficulty to export samples from West Africa to Europe, as well as to organize a unique collection, the “centralized” Biobank foreseen by the project has to be considered as a “disseminated” Biobank. It includes the different BioBanks in the several labs and institutes, partners of the project. This gathers all the samples stored in the several field laboratories and institutes involved in the project, and these samples can be used to carry out the validation tests in the same laboratory where the samples are stored or, if possible, by shipping the samples among the partner laboratories.

Capacity building and knowledge transfer are just a few major factors which will contribute to successful preparedness strategies to face the new Ebola epidemics. The WP5 have contributed to build several platforms for the responsible entities and clinical staff in West Africa in terms of field diagnostics, and clinical aspects of the Ebola virus disease by organizing workshops. During the second period, we have initiated a knowledge exchange platform within the mobile laboratories from European mobile lab, PHE, Pasteur Institute, INMI, Nigerian, South African, Dutch, and US CDC mobile labs, by organizing a workshop by in Dakar, in February 2016.

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