Fta whatman paper

Blood from finger pricks was collected on filter paper (FTA^® Whatman paper) on day 0 and the follow-up days, dried and stored in sealed plastic bags. Samples with positive asexual parasitaemia on/or after the seventh day were subjected for molecular analysis. Briefly, DNA was extracted from day 0 and the respective days of re-appearance of parasites using the QIAamp^® DNA Mini Kit (Qiagen, Hilden, Germany) for dried blood according to manufacturer protocol. Nested PCR (nPCR) was performed for identification of P. falciparum MSP1 and MSP2 allele variants, as described previously [16 (link)]. The results were indicated as recrudescence, re-infection or unknown for two lost samples according to WHO guidelines [17 ].

Guardians or parents were requested to return with the children for follow-up. Blood from finger pricks was collected on filter paper (FTA®Whatman paper) on day 0,1,2,3,7,14,21,28,35 for PCR genotyping of Merozoite Surface Protein 1(MSP1) and Merozoite Surface Protein (MSP2) to distinguish between recrudescence and reinfection, MSP1 and MSP2 alleles to establish genetic diversity and allele frequencies (to be published elsewhere) and CYP3A4*1B plus CYP3A5*3 to establish association with plasma concentrations. Venous blood (2mls) was also collected on day 0,1,2,3,7 and 14 for lumefantrine and piperaquine quantification (to be published elsewhere). Thick and thin blood smears were stained by Giemsa (on each day of the visit) according to the WHO standard protocol [23 ]. Parasite identification and counting were done by two independent experienced microscopists.