A mass spectrometry assay for quantitative analysis of P. falciparum Histidine Rich Protein 2
Significant variation has been reported in the field performance of rapid diagnostic tests (RDTs) to accurately diagnose malaria infections via the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP2).
An important variable that may influence test performance is the presence of significant sequence polymorphism in PfHRP2 sequences from geographically distinct P. falciparum isolates, resulting in variable recognition from the monoclonal antibodies (MABs) used in the RDTs. Such limitations (i.e., variable reactivity, cross-recognition e.g., monoclonal antibodies against PfHRP2 cross-reacting with PfHRP3, and non-specific binding) are well recognized fundamental challenges to the utility of conventional competitive or sandwich immunoassay based detection strategies.
Here, a quantitative mass spectrometry based assay that couples targeted protein capture with stable isotope labelled peptide internal standards, protein digestion, high-performance liquid chromatography and tandem mass spectrometry (LC-MS/MS) detection, is being developed and utilized to identify and quantify peptides specific to PfHRP2 and its individual sequence variants for in vitro clinical diagnostic analysis in human plasma samples infected with P. falciparum. This research is being performed as part of a new collaboration with Professor James McCarthy at the Queensland Institute of Medical Research.
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