Development of a CRISPR-Cas12-based isotheral PCR for the detection of parasitic diseases in an easy customizable lateral flow format, targeting T. evansi and T. equiperdum Trypanosomosis

Animal Trypanosomosis is a parasitic infection that threatens livestock worldwide, including in
Europe. Due the absence of an effective vaccine approach, the only control measures available are
accurate diagnosis followed by treatment. In Europe in particular, a positive diagnosis for T. evansi or
T. equiperdum is obligatory followed by slaughter of the animal as treatment is prohibited.
Unfortunately, today methods for trypanosome identification lack uniformity. Moreover, antibodybased
testing methods suffer from low positive predictive value. As T. evansi poses a zoonosic risk, a
One Health Approach is required to limit its impact. Hence, simple-to-use diagnostics, which can be
implemented at a point- of-care (POC), are needed. These tools should excel by their high positive
and negative predictive value, combined with a high level of sensitivity. Hence, we propose to design
a CRISPR-Cas12 LAMP-based lateral flow test, capable of detecting T. evansi and the closely related
T. equiperdum, by using an isothermal amplification approach linked to the CRISPR-Cas12 technology
and performing a lateral flow readout. By developing these tests, the assay can be directly
implemented into control strategies to tackle trypanosomosis on a global scale and can replace the
cumbersome and low-sensitive microscopy screening that is still the main screening method today.
Such test can also replace the low-accuracy antibody-tests that cannot distinguish between active
and past infection.