Improving Molecular Epidemiological Surveillance of Strongyloidiasis Upon Differentiation of <i>Strongyloides fuelleborni fuelleborni</i> From <i>Strongyloides stercoralis</i>

Molecular epidemiological surveillance for zoonotic strongyloidiasis is confounded by a genus-specific TaqMan probe assay that conflates Strongyloides fuelleborni fuelleborni with Strongyloides stercoralis. To improve surveillance, we developed and validated a novel duplex species-specific TaqMan probe assay, screening a representative collection of available clinical samples. Our assay was highly discriminatory, evidencing no cross-reactivity, and had a lowest DNA detection limit of 1 pg/µL. However, as the genus-specific DNA assay has greater detection ability, we propose a 2-step protocol where samples are first screened with this assay then, if positive, screened with our species-specific assay, discriminating (co)infections between each threadworm species.