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The first imported case of Plasmodium knowlesi in Scotland is described in a 33-year-old female with a travel history to Borneo. The patient ceased to take antimalarial prophylaxis after 4 days of her 10-day visit and presented with a history of fever, rigor, vomiting, and diarrhea after 13 days on her return to the UK. Malaria antigen detection using the Optimal-IT and Binax-NOW kits was negative. Unusual trophozoite-like structures were observed under microscopic examination and the identification of P. knowlesi performed by a nested polymerase chain reaction (PCR) gel-based approach was confirmed by using a PCR-sequencing assay.