Laboratory challenges of Plasmodium species identification in Aceh Province, Indonesia, a malaria elimination setting with newly discovered P. knowlesi

摘要

Author summary In Southeast Asia, Plasmodium knowlesi, a malaria parasite of macaques, was recently discovered to infect humans. This emerging disease is important because it has potential for causing severe disease and death, and it is a threat to malaria elimination efforts in the region. In this report from Aceh Province, Indonesia, where P. knowlesi was only recently discovered, the authors report on the laboratory challenges of distinguishing this species from other human species. Using several different molecular methods, they investigated 41 malaria cases which by microscopy, were initially reported as: P. falciparum (49%), P. vivax (39%), P. malariae (7%), and indeterminate (5%). Only after using a P. knowlesi-specific nPCR method and sequencing, did they find that nearly half were P. knowlesi. Consistent with a sparse literature, a field-friendly molecular method (genus-specific LAMP) reliably detected P. knowlesi, while use of a more standard reference laboratory molecular method (18S rRNA nPCR targeting the four human-only species) missed the infections. Also another reference laboratory molecular method (cytb nPCR) mis-classified P. knowlesi infections as P. vivax due to cross-reactivity. To address the emerging threat of P. knowlesi, new detection methods are needed for point-of-care and reference testing.