PROGRAM PROJECT 3 [PP3]: Advancing Clinical and Entomological Surveillance (ACES) through emergent diagnostic and translational systems biology platforms to address emerging issues viz. asymptomatic malaria and non-falciparum malaria transmission in the context of malaria elimination and eradication (MEE). ABSTRACT/SUMMARY Progress toward the global target set out in the WHO Global Technical Strategy for malaria 2016-2030 to reduce malaria case incidence and mortality rates by at least 90% by 2030― against a 2015 baseline— has stalled. A significant challenge to effectively identifying and treating malaria infection is targeting asymptomatic cases – important parasite reservoirs contributing to sustained transmission. Substantial emphasis needs to be placed on identifying subclinical P. falciparum (Pf) infection to reach global elimination goals, especially considering difficulties that are arising from increasing failure rates of rapid diagnostic tests (RDT) – and thus increasing false-positive rates – for PfHRP2-based RDTs due to strains with pfhrp2/3 gene deletions, as well as a considerable lack of sensitive field-deployable diagnostic tools for non-falciparum malaria (NFM), which includes P. vivax (Pv), P. ovale curtisi (Poc), P. ovale wallikeri (Pow), and P. malariae (Pm). To better understand the epidemiological factors contributing to subclinical Pf and/or NFM infections and their role in residual malaria transmission, Plasmodium species detection is paramount, as it can influence treatment regimens and potentially guide control and intervention policy. While malaria RDTs and the ‘gold standard’ light microscopy remain important, the requirement to quantify the subclinical parasite reservoir alongside the detection of key mutations necessitates the use of molecular methods. Going beyond the molecular approach, other ‘omics’ approaches may be more powerful in providing new insight into malaria transmission biology. The overarching hypothesis to be explored in PP3 is to investigate the use of innovative tailored multiomics solutions to address the considerable challenge of accurately detecting and diagnosing subclinical Pf and NFM infections. Leveraging our existing bank of saliva, whole blood, dried blood spots, and liquid blood spots, the project will seek to test, define, and validate a suite of novel diagnostic tests to detect, quantify and characterize asymptomatic Pf infection and NFM. T his Émergents ICEMR program project seeks to: Aim PP3.1. Implementation of non- invasive rapid tests for identifying individuals (adults and children) and thereby quantifying the parasite reservoir in the human population contributing to residual transmission; Aim PP3.2. Developing and validating a quantitative dried blood spot into direct, no extraction qPCR diagnostic platform for Pf, Pv, Po(w), Po(c), and Pm; and Aim PP.3.3. Dissecting the biology of asymptomatic Pf, NFM and mixed infections viz. malaria transmission. Key outcomes of PP3 will be to...