Fig. 2

Methodological flow scheme. Microscopy diagnosis was confirmed by nested multiplex PCR, distinguishing P. falciparum, Plasmodium vivax, Plasmodium Ovale, and Plasmodium malaria. Four independent PCRs were run for the P. falciparum samples to detect deletions in exon 1–2 and exon 2 of pfhrp2 and pfhrp3. The deletion of any exon was confirmed by an absence of amplification after three PCR repetitions and the confirmation of maintained DNA quality. Samples that could not be confirmed for DNA quality were excluded from the analysis for that gene. Subsequent data analysis for the combined results was performed only with those samples included in the independent analysis for both genes. Additionally, a sub-sample lacking the deletion for exon 2 of both genes was sequenced and the genetic diversity and variation in amino-acid sequences analysed. Finally, a sub-sample of these sequences was used to predict the protein structure model and to locate the epitopes in the structure