Dried Blood Spot Extraction Protocol

Dried blood spot extraction was adapted from a previously reported method.^{16 (link)} Dried blood spot was prepared by depositing 10 μL of parasitized whole blood onto Whatman 903 protein saver cards. The spots were allowed to air-dry for a minimum of 4 hours, removed using a 6-mm biopsy punch, and placed in 2-mL microcentrifuge tubes (one spot per tube). Next, 200 μL of phosphate-buffered saline (PBS) with 0.1% Tween-20 (PBST) was added to each tube. The tubes were vortexed at 3,200 rpm for 10 minutes and then placed in a mini-centrifuge for 30–60 seconds. The supernatant was removed and saved for analysis.