Molecular Identification of Plasmodium Species

The dried blood spots were sent to the Malaria Research Centre, University of Malaysia, Sarawak, Malaysia, to identify the Plasmodium species through molecular analysis. The DNA was extracted from the dried blood spots using InstaGene (Bio-Rad Laboratories, USA), as described previously [46 (link)]. The DNA samples were initially analyzed through a nested polymerase chain reaction (PCR) with genus-specific primers for detecting Plasmodium, as described previously [47 (link)]. The Plasmodium-positive samples were then analyzed through a nested PCR with species-specific primers for P. knowlesi, P. coatney, P. cynomolgi, P. inui, and P. fieldi parasites, as described previously [48 (link)].