Project2: Developing New Diagnostic Tools for the Detection of Plasmodium Infection Across the Spectrum of Disease Severity

PROJECT 2 SUMMARY Novel and more granular methods of diagnosis of Plasmodium infection are urgently needed in India, where several important barriers to elimination have recently come to light. Through the development of new and innovative detection tools across the spectrum of disease severity, Project 2 and its three subprojects broadly aim to support and further India’s efforts toward malaria elimination. To help tackle the high prevalence of Pfhrp2/ Pfhrp3 deletions reported in Odisha, Subproject 1 will identify novel RDT biomarker candidates by deep learning techniques to mine the P. falciparum genome and proteome. These potential new biomarkers will be filtered for relative proteomic abundance and specific characteristics such as chromosomal location and essentiality, and protein binders designed against them. Selected designed protein binders will be immobilized on a nitrocellulose test strip and tested at our Indian field sites for sensitivity and specificity and benchmarked against PfHRP2 rapid detection tests commonly used in India. Subproject 2 will evaluate ophthalmologic and plasma biomarker signatures to identify patients at risk of cerebral and cognitive changes following falciparum malaria. The prevalence of short and long-term neuro-disabilities in adults recovering from falciparum malaria will be assessed using serial quantitative magnetic resonance imaging methods, ophthalmologic and neuropsychometric assessments, permitting the detection of a potentially considerable but unrecognized impact of Plasmodium infection on brain function not limited to WHO-defined cerebral malaria patients and inform follow-up and rehabilitation strategies. Lastly, Subproject 3 will combine plasma RNA sequencing with proteomic screening to determine circulating biomarkers associated with the presence of cryptic malaria parasites in the spleen of individuals with asymptomatic infection. Circulating miRNA and protein profiles in blood samples from patients undergoing traumatic splenectomy and categorized as with or without splenic parasites in Project 1 will be compared. Candidate biomarkers associated with splenic carriers of Plasmodium parasites will be tested at our three field sites to assess the prevalence of this reservoir in Odisha. Identifying, developing, and validating new biomarkers across the whole spectrum of Plasmodium infection will provide valuable tools towards diagnosis, prognosis, and treatment of infected individuals, with the overarching goal to support India's efforts towards malaria elimination both at state and national levels.