Frozen blood clots can be used for the diagnosis of distinct Plasmodium species in man and non-human primates from the Brazilian Atlantic Forest

Table 1 PCR results from whole blood and blood clots according to malaria species and estimated parasitaemia according to microscopy examination (thick and thin blood film)

Code	Species	Parasitaemia (parasites/μL)	PCR^a: whole blood	PCR^a: 1st blood clot DNA extraction^b	PCR^a: 2nd blood clot DNA extraction^b	PCR^a: 3rd blood clot DNA extraction^b
H1	Homo sapiens	208	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium
H2	Homo sapiens	368	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium	Plasmodium; P. vivax/P. simium
H3	Homo sapiens	0	−	−	−	−
M1	Alouatta g. clamitans	300	Plasmodium; P. vivax/P. simium and P. malariae/P. brasilianum	Plasmodium; P. vivax/P. simium and P. malariae/P. brasilianum	Plasmodium; P. vivax/P. simium and P. malariae/P. brasilianum	Plasmodium; P. vivax/P. simium and P. malariae/P. brasilianum
M2	Callithrix hybrid	0	−	−	−	−
M3	Alouatta g. clamitans	40	P. malariae/P. brasilianum	P. malariae/P. brasilianum	P. malariae/P. brasilianum	P. malariae/P. brasilianum

^aGene targets were cysteine proteinase for P. vivax, ssrRNA for P. malariae and 18S rRNA for genus Plasmodium; ^b DNA extractions from clots disrupted by high-speed shaking (6000 rpm/40 s) in lysis buffer, without glass beads; −: negative result

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