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The simian parasite Plasmodium knowlesi is now the commonest cause of malaria in Malaysia and can rapidly cause severe and fatal malaria. However, microscopic misdiagnosis of Plasmodium species is common, rapid antigen detection tests remain insufficiently sensitive and confirmation of P. knowlesi requires polymerase chain reaction (PCR). Thus available point-of-care diagnostic tests are inadequate. This study reports the development of a simple, sensitive, colorimetric, high-throughput loop-mediated isothermal amplification assay (HtLAMP) diagnostic test using novel primers for the detection of P. knowlesi. This assay is able to detect 0.2 parasites/μL, and compared with PCR has a sensitivity of 96% for the detection of P. knowlesi, making it a potentially field-applicable point-of-care diagnostic tool.
Financial support: Malaysian Ministry of Health (Grant no. BP00500420), AusAlD Asia-Pacific Malaria Elimination Network (Grant no. 108-07), Australian National Health and Medical Research Council (Program grant no. 1037304, Project Grant no. 1045156, Practitioner fellowship to NMA no. 1042072 and JSM no. 1041802, scholarship to SB no. 1055410 and MJG no. 1074795) and Queensland Health Research Fellowship to JSM.
Authors' addresses: Sumudu Britton and James S. McCarthy, Clinical Tropical Medicine, QIMR Berghofer Medical Research Institute, Brisbane, Australia, and School of Medicine, University of Queensland, Brisbane, Australia, E-mails: firstname.lastname@example.org and email@example.com. Qin Cheng, Department of Drug Resistance and Diagnostics, Australian Army Malaria Institute, Brisbane, Australia, E-mail: firstname.lastname@example.org. Matthew J. Grigg and Nick M. Anstey, Global Health Division, Menzies School of Health Research, Charles Darwin University, Darwin, Australia, E-mails: email@example.com and firstname.lastname@example.org. Timothy William, Clinical Research Center, Queen Elizabeth Hospital, Kota Kinabalu, Sabah, Malaysia, and Infectious Diseases, Jesselton Medical Centre, Kota Kinabalu, Sabah, Malaysia, E-mail: email@example.com.