A specific, stable, and accessible LAMP assay targeting the HSP70 gene of Trypanosoma cruzi
Guarnizo, S. A. G.; Basma, L.; Equila, S.; Condori, B. J.; Malaga, E.; Defazio, S.; Arteaga, E.; Valverde, J. K.; Obregon, M.; Takyar, A.; Duque, C.; Hakim, J.; Tinajeros, F.; Gilman, R.; Bowman, N.; Mugnier, M.
Show abstract
Diagnostic delays prevent most Chagas disease patients from receiving timely therapy during the acute phase when treatment is effective. qPCR-based diagnostic methods provide high sensitivity during this phase but require specialized equipment and complex protocols. More simple and cost-effective tools are urgently needed to optimize early Chagas disease diagnosis in low-income endemic regions. Here, we present a loop-mediated isothermal amplification (LAMP) that targets a highly conserved region in the HSP70 gene of Trypanosoma cruzi, the causative agent of Chagas disease. This assay demonstrates species-specific amplification across multiple parasite genetic lineages while maintaining stability after 2 hours of incubation and at least 8 months of storage at -20{degrees}C. Moreover, the assay is at least 12 times less expensive than the TaqMan qPCR that is currently routinely used for acute Chagas diagnostics. Population-based validation in 100 infants born to Chagas-positive mothers in Santa Cruz, Bolivia, yielded a specificity of 100% and sensitivity exceeding 77% when compared to a TaqMan qPCR that targets satellite DNA. This cost-effective assay holds promise for large-scale diagnosis of Chagas disease in endemic regions with limited resources.
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