Artemether-lumefantrine is one of the first-line treatments for uncomplicated malaria, however, it is threatened by resistance. Four resistance markers towards lumefantrine (LMF) are Plasmodium falciparum multidrug resistance 1 (pfmdr1) N86, 184F and D1246 and Plasmodium falciparum chloroquine resistance transporter (pfcrt) K76. WHO emphasises the need for monitoring of resistance markers as one of the most important interventions to combat resistance, and two potential tools for this are wastewater surveillance and xenomonitoring. Plasmodium spp. DNA has been detected in wastewater and hospital effluents, indicating that wastewater surveillance could be used, even if the parasite is not transmitted by human waste, but rather by vectors. Hyperplex PCR™ can detect polymorphisms on a single-nucleotide level, allowing for surveillance of molecular markers of resistance without the need for sequencing. This study evaluated wastewater (n=6) and Anopheles mosquitoes (n=200), as non-clinical matrices for detection of Plasmodium spp. and LMF resistance markers in Uganda, using qPCR. No detection of Plasmodium spp. in wastewater samples was found. The 22 parasite-positive mosquitoes pools were further analysed with Hyperplex PCR™ for LMF resistance markers. LMF resistance markers were observed in a majority, 77.5% for N86, 96.4% for 184F and 100% for D1246 and K76 in Anopheles mosquitos. No definitive conclusion can be made regarding wastewater surveillance of Plasmodium spp., due to the small sample size and relatively low malaria prevalence setting and season. Nonetheless, xenomonitoring in combination with Hyperplex PCR™ demonstrates potential as an antimalarial resistance marker surveillance methodology.