Periodic Reporting for period 2 - FILODIAG (Ultra-Fast Molecular Filovirus Diagnostics - Sofia ref.: 115844)
Reporting period: 2016-01-01 to 2017-12-31
The aim of this project was to develop an ultra-fast and accurate molecular testing platform including an instrument, assay and workflow that can perform a molecular Ebola virus test within 10-15 minutes. In emergency situations, the promptly isolation and treatment of Ebola-infected patients is a critical need at treatments centers to content the transmission and to control deadly outbreaks. Patients, suspected of infection with Ebola, due to fever and/or other clinical signs, could be rapidly and accurately tested even at resource-limited settings.
During the project, the Pharos400 prototype instrument and software were redesigned and optimized to allow for an integrated reverse-transcription step leading to the one-step RT-Laser PCR. Moreover, the Pharos400 was designed to be much more robust for the field, and the cartridges were improved for better handling. The detection method of the Laser PCR system was completely re-engineered, and eventually comprised specific probes for improved test accuracy.
GNA developed a protocol for the lyophilisation of the reagents to enable a ready-to-use Laser PCR mix with a shelf-life of 2 months at room temperature, or even longer when cooled. A one-step RT-Laser PCR duplex assay for the Ebola + IC (internal process control) targets was developed and optimized by GNA, and successfully tested by INMI and partners in the field in Sierra Leone on ribonucleic acids isolated from diverse samples (plasma, urine, and oral), derived from Ebola-infected patients from the 2014-2015 outbreak.
The Laser PCR technology, subject to this project, is now already commercially available through GNA Biosolutions GmbH. The experiences with the Pharos400 prototype, with its Laser PCR amplification and fluorescence-based detection, laid the foundation for developing and launching an industrialized laboratory instrument, the Pharos V8, after this project.
Up to now, the project has already generated a patent application, articles, and posters at international scientific meetings.
GNA developed a protocol for the lyophilisation of the reagents to enable a ready-to-use Laser PCR mix with a shelf-life of 2 months at room temperature, or even longer when cooled. A one-step RT-Laser PCR duplex assay for the Ebola + IC (internal process control) targets was developed and optimized by GNA, and successfully tested by INMI and partners in the field in Sierra Leone on ribonucleic acids isolated from diverse samples (plasma, urine, and oral), derived from Ebola-infected patients from the 2014-2015 outbreak.
The Laser PCR technology, subject to this project, is now already commercially available through GNA Biosolutions GmbH. The experiences with the Pharos400 prototype, with its Laser PCR amplification and fluorescence-based detection, laid the foundation for developing and launching an industrialized laboratory instrument, the Pharos V8, after this project.
Up to now, the project has already generated a patent application, articles, and posters at international scientific meetings.
The ultra-fast detection of the Ebola virus with Laser PCR has been successfully demonstrated. It was shown that a much faster PCR based assay can be implemented, which provides comparable results after a few minutes as compared to conventional methods which typically require 40-60 minutes. Even though at this stage the full workflow has not been integrated yet, the successful proof-of-concept paves the way for a new generation of molecular diagnostics at the Point-of-Need, providing screening opportunities which can be crucial for containing further outbreaks. Such fast Point-of-Need screening can be pivotal for earlier treatment as patients can wait for their test results and do not need to be tracked.