During the first year, the team discussed pros and cons of PoC Lyme tests with potential end-users. The conclusion was that a true PoC test would not be needed, but that at least the first step should be close to point of blood drawing. We performed a Health Technology Assessment to quantify the economic burden and benefits for patients and society and looked at reimbursement schemes in The Netherlands, Sweden, Germany and Austria.
Technically, a vast amount of work was done to design and develop a device (named WOLF) that would enable performing the stimulation step of the test at the point of blood drawing and thus help overcome the logistic hurdles for CMI tests. We further investigated the optimal Borrelia antigen mixture to be used for stimulation step and assessed stability and the storage conditions. Antigen production at DIARECT was optimized to enable kit manufacturing once all components of the kit are ready. For read-out of the relevant CMI biomarkers (cytokines), the consortium compared sensitivity and cost of three methods, in order to make a sensitive test affordable to many people.
In the second year, the team came to the conclusion that despite extensive troubleshooting efforts at Innatoss, the Lyme CMI-test was not fit to distinguish between infected and non-infected individuals. The same was found upon testing other marketed LB CMI-tests for LB . Therefore, efforts to develop a novel Lyme-CMI test were abandoned. This was important information for the Lyme community since CMI-tests for Lyme are often claimed to outperform the classical serological tests.
The WOLF device developed in period 1, however, was considered to be an immensely useful tool to open up CMI-based diagnostics in any disease context. Therefore, the team continued this effort. The devices were manufactured in China, directed by npk design. A patent application was filed in 2017 and is under review.
In the 3rd period, Innatoss completed development of two sensitive, enzyme-linked immunosorbent assays (ELISA) for measuring cytokines to be used in the second, read-out step of CMI-tests. All relevant parameters of assay performance, including stability, precision, interference, and sensitivity were validated. Various redesigns of the WOLF devices were produced and tested. Risk analyses were performed and release procedures written. The team also explored other applications of both the ELISAs and the WOLF device.
During the final sprint towards the end of the project, COVID-19 changed the world around us. The COVID-19 pandemic thwarted any planning, first in China, and later in Europe. Production of devices in China was delayed, clinical studies became virtually impossible, due to the fact that hospitals closed and health care professionals were hard to find. As a team, we dealt in a positive manner with the continuing changes in planning. The Commission facilitated it by extending the program.
Our manufacturer in China re-opened its facilities quickly, continuing with required changes. The validation lots of the ELISAs were produced just before the world came to a halt, and testing performance continued during the pandemic. User reviews and stakeholder meetings could not be organized, but the team creatively got the feedback it needed: A group of medical students joined Innatoss for COVID-19 testing. Being future users, they were asked to provide feedback on the use of the devices.
In Vienna, the application of WOLF for Chlamydia was slowed down due to closure of the out-patient clinic. Nevertheless, the team made substantial progress in generating materials for a future test.
