Background Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) is widely used for sand fly identification, but its potential to detect Leishmania infections in vectors remain underexplored. This pilot study evaluated whether MALDI-ToF MS protein profiles of lab-reared Lutzomyia longipalpis and Nyssomyia neivai can discriminate Leishmania infantum-infected from uninfected females. Methodology Colonies were experimentally infected with L. infantum using membrane feeding, and females were collected at different days post-blood meal. Thoraces and legs were processed individually for MALDI-ToF MS, and spectra were analysed using both Bruker software and custom R pipelines. Principal findings Unsupervised approaches (MSP dendrograms, PCA) showed limited or inconsistent separation of infection status for Lu. longipalpis. In contrast, supervised machine-learning models built on peak-intensity matrices achieved excellent discrimination between infected and uninfected specimens for both species, with several algorithms reaching near-perfect performance on an external test set not used for training. Variable-importance analysis highlighted sets of m/z peaks, mainly showing decreased intensity in infected sand flies, as putative infection biomarkers. Conclusion This proof-of-concept study highlights that L. infantum infection induces reproducible, species-specific alterations in sand-fly MALDI-TOF profiles, supporting further development of high-throughput, MS-based screening of infected vectors.
de Souza, L. A. F., Kariya, E., Prudhomme, J., Depaquit, J., Vieira da Costa-Ribeiro, M. C., Huguenin, A.
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