Mwesigwa, AlexGolumbeanu, MonicaJones, SamCantoreggi, Sara L.Musinguzi, BensonNankabirwa, Joaniter I.Bikaitwoha, Everd ManipleKalyango, Joan NKaramagi, CharlesPlucinski, MateuszNsobya, Samuel L.Nsanzabana, ChristianByakika-Kibwika, Pauline2025-02-062025-02-062025-02-05Mwesigwa, A., Golumbeanu, M., Jones, S., Cantoreggi, S. L., Musinguzi, B., Nankabirwa, J. I., ... & Byakika-Kibwika, P. (2024). Assessment of different genotyping markers and algorithms for distinguishing Plasmodium falciparum recrudescence from reinfection in Uganda.https://dir.muni.ac.ug/handle/20.500.12260/722Antimalarial therapeutic efficacy studies are vital for monitoring the efficacy of antimalarial drugs in malaria-endemic regions. The WHO recommends genotyping of polymorphic markers including msp-1, msp-2, and glurp to aid distinguishing recrudescences from reinfections. Recently, WHO proposed replacing glurp with microsatellites (Poly-α, PfPK2, TA1). However, suitable combinations with msp-1 and msp-2 have not been systematically assessed. Additionally, the performance of different algorithms for classifying recrudescence is unclear. This study investigated various microsatellites alongside msp-1 and msp-2 for molecular correction and compared genotyping algorithms across three malaria-endemic areas in Uganda. Microsatellites 313, Poly-α, and 383 exhibited the highest diversity, while PfPK2 and Poly-α revealed elevated multiplicities of infection across all sites. The 3/3 match-counting algorithm classified fewer recrudescences than the ≥ 2/3, and Bayesian algorithms at both ≥ 0.7 and ≥ 0.8 probability cutoffs. The msp-1/msp-2/2490 combination identified more recrudescences using the ≥ 2/3 and 3/3 algorithms in the artemether-lumefantrine (AL) treatment arm, while msp-1/msp-2/glurp combination identified more cases of recrudescence using the ≥ 2/3 in the dihydroartemisinin-piperaquine (DP) arm. Microsatellites PfPK2 and Poly-α, potentially sensitive to detecting minority clones, are promising replacements for glurp. Discrepancies in recrudescence classification between match-counting and Bayesian algorithms highlight the need for standardized PCR correction practices.Biological sciences/Molecular biologyHealth sciences/Health careHealth sciences/Medical researchPlasmodium falciparumRecrudescenceReinfectionMicrosatellitesMsp-1Msp-2Antimalarial drug