Grabias B, et al. - As microscopy is time and labour-intensive and can only reach typical detection limits of 5000–150,000 parasites/mL of blood, researchers here sought for highly sensitive molecular methods for detection of low-grade sub-microscopic infections for surveillance studies and identifying asymptomatic reservoirs of malaria transmission. They developed and characterized PCR-based nucleic acid assays utilizing primers specific to the P. falciparum 18S ribosomal gene sequence and highly conserved regions of the abundant (approximately 60 variants) P. falciparum var. gene EMP1 in this report. As per data from analytical studies, using the PfEMP1 primer set for P. falciparum detection consistently outperforms the traditional 18S primer set, detecting as little as 0.01 pg of Plasmodium gDNA and 9.3 parasites/mL compared to 0.07 pg and 98.2 parasites/mL for the Pf18S primer set. Results thereby establish that for highly sensitive detection of both acute infections from filter paper samples and submicroscopic asymptomatic low-grade infections, PfEMP1 is a novel amplification target.