Fig. 1

Flowchart of PCR techniques and alignment of the Plasmodium cytochrome oxidase gene III PCR products. For the cytochrome oxidase III (COX-III) gene the expected sizes of PCR fragments are: P. vivax (506 bp), P. falciparum (508 bp), P. malariae (504 bp), P. knowlesi (499 bp), P. ovale wallikeri and P. ovale curtisi (506 bp). With the COX-III gene sequence alignment, 140 polymorphisms (28 % of nucleotides) provide information for species diagnosis (there are eight blocks of highly polymorphic regions). For differentiation between P. ovale wallikeri and P. ovale curtisi, 16 SNPs were found (green arrows). When using 18s-rRNA nested PCR, additional Nest2 PCRs with species-specific primers are required for diagnosis of P. malariae, P. ovale wallikeri, P. ovale curtisi and P. knowlesi