Characterization of plasmodium lactate dehydrogenase and histidine-rich protein 2 clearance patterns via rapid on-bead detection from a single dried blood spot

Christine F. Markwalter, Vanderbilt University
Lauren E. Gibson, Vanderbilt University
Lwiindi Mudenda, Vanderbilt University
Danielle W. Kimmel, Vanderbilt University
Saidon Mbambara, Macha Research Trust
Philip E. Thuma, Macha Research Trust
David W. Wright, Vanderbilt University


A rapid, on-bead enzyme-linked immunosorbent assay for Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein 2 (HRP2) was adapted for use with dried blood spot (DBS) samples. This assay detected both biomarkers from a single DBS sample with only 45 minutes of total incubation time and detection limits of 600 ± 500 pM (pLDH) and 69 ± 30 pM (HRP2), corresponding to 150 and 24 parasites/µL, respectively. This sensitive and reproducible on-bead detection method was used to quantify pLDH and HRP2 in patient DBS samples from rural Zambia collected at multiple time points after treatment. Biomarker clearance patterns relative to parasite clearance were determined; pLDH clearance followed closely with parasite clearance, whereas most patients maintained detectable levels of HRP2 for 35–52 days after treatment. Furthermore, weak-to-moderate correlations between biomarker concentration and parasite densities were found for both biomarkers. This work demonstrates the utility of the developed assay for epidemiological study and surveillance of malaria.