A next generation CRISPR diagnostic tool to survey drug resistance in Human African Trypanosomiasis

The WHO aims to eliminate the gambiense form of human African trypanosomiasis (HAT) by 2030. With the decline of reported cases, maintaining efficient epidemiological surveillance is essential, including the emergence of drug-resistant strains. We have developed new highly specific diagnostic tools using Specific High-Sensitivity Reporter Enzymatic UnLOCKing (SHERLOCK) technology for monitoring the presence of drug-resistant genotypes that (1) are already circulating, such as the AQP2/3 ₍₈₁₄₎ chimera providing resistance to pentamidine and melarsoprol, or (2) could emerge, such as Tb CPSF3 (N ²³² H), associated to acoziborole resistance in lab conditions. The melarsoprol - pentamidine AQP2/3 ₍₈₁₄₎ SHERLOCK assay detected RNA from both cultured parasites and field isolated strains from gHAT patients in relapse following treatment. The acoziborole CPSF3 _(SNV) SHERLOCK assay discriminated between wild-type CPSF3 RNA and CPSF3 with a single A-C mutation that confers resistance to acoziborole in vitro .