Malpartida-Cardenas, K.Moser, N.Ansah, F.Pennisi, I.Prah, D.A.Amoah, L.E.Awandare, G.Hafalla, J.C.R.Cunnington, A.Baum, J.Rodriguez-Manzano, J.Georgiou, P.2023-08-142023-08-142023http://ugspace.ug.edu.gh:8080/handle/123456789/39751Research ArticleHuman malaria is a life-threatening parasitic disease with high impact in the sub-Saharan Africa region, where 95% of global cases occurred in 2021. While most malaria diagnostic tools are focused on Plasmodium falciparum, there is a current lack of testing non-P. falciparum cases, which may be underreported and, if undiagnosed or untreated, may lead to severe consequences. In this work, seven species-specific loop-mediated isothermal amplification (LAMP) assays were designed and evaluated against TaqMan quan titative PCR (qPCR), microscopy, and enzyme-linked immunosorbent assays (ELISAs). Their clinical performance was assessed with a cohort of 164 samples of symptomatic and asymptomatic patients from Ghana. All asymptomatic samples with a parasite load above 80 genomic DNA (gDNA) copies per mL of extracted sample were detected with the Plasmodium falciparum LAMP assay, reporting 95.6% (95% confidence interval [95% CI] of 89.9 to 98.5) sensitivity and 100% (95% CI of 87.2 to 100) specificity. This assay showed higher sensitivity than microscopy and ELISA, which were 52.7% (95% CI of 39.7 to 67%) and 67.3% (95% CI of 53.3 to 79.3%), respectively. Nine samples were posi tive for P. malariae, indicating coinfections with P. falciparum, which represented 5.5% of the tested population. No samples were detected as positive for P. vivax, P. ovale, P. knowl esi, or P. cynomolgi by any method. Furthermore, translation to the point-of-care was demonstrated with a subcohort of 18 samples tested locally in Ghana using our hand held lab-on-chip platform, Lacewing, showing comparable results to a conventional fluo rescence-based instrument. The developed molecular diagnostic test could detect asymp tomatic malaria cases, including submicroscopic parasitemia, and it has the potential to be used for point-of-care applicationsennucleic acid amplification,malariadiagnosticspoint-of-careArticle