Development, validation and evaluation of a rapid PCR-nucleic acid lateral flow immuno-assay for the detection of Plasmodium and the differentiation between Plasmodium falciparum and Plasmodium vivax

Table 1 Overview of the primer sequences and used concentrations in the PCR-NALFIA reaction

Target	Primer type	Sequence	Product size (bp)	Concentration in PCR mix (nM)
GAPDH	Forward	Biotin-5’ TGCACCACCAACTGCTTAGC -3’	90	75
GAPDH	Reverse	Texas Red-5’ GGCATGGACTGTGGTCATGAG -3’		75
Pan-Plasmodium	Forward	Digoxigenin-5’ TCAGATACCGTCGTAATCTTA -3’	180	50
Pan-Plasmodium	Reverse *	Biotin-5’- AACTTTCTCGCTTGCGCG -3’		900
P. falciparum	Forward	FAM-5’ GTCATCTTTCGAGGTGACTT -3’	100	200
P. vivax	Forward	DNP-5’ TTTCTCTTCGGAGTTTATTC -3’	100	650

* This primer was used as a reverse primer for the P. falciparum and P. vivax reaction as well.