Journal
JOURNAL OF INFECTIOUS DISEASES
Volume 208, Issue 4, Pages 645-652Publisher
OXFORD UNIV PRESS INC
DOI: 10.1093/infdis/jit184
Keywords
malaria; diagnosis; LAMP; loop-mediated isothermal amplification; sensitivity and specificity; PCR; polymerase chain reaction; molecular diagnosis; DNA; Plasmodium falciparum; Uganda; Africa
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Funding
- Foundation for Innovative New Diagnostics through Bill and Melinda Gates Foundation
- Ministry of Foreign Affairs of the Government of the Netherlands
- United Kingdom Department for International Development
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Background. Current malaria diagnostic tests, including microscopy and antigen-detecting rapid tests, cannot reliably detect low-density infections. Molecular methods such as polymerase chain reaction (PCR) are highly sensitive but remain too complex for field deployment. A new commercial molecular assay based on loop-mediated isothermal amplification (LAMP) was assessed for field use. Methods. Malaria LAMP (Eiken Chemical, Japan) was evaluated for samples from 272 outpatients at a rural Ugandan clinic and compared with expert microscopy, nested PCR, and quantitative PCR (qPCR). Two technicians performed the assay after 3 days of training, using 2 alternative blood sample-preparation methods and visual interpretation of results by fluorescence assay. Results. Compared with 3-well nested PCR, the sensitivity of both LAMP and single-well nested PCR was 90%; the microscopy sensitivity was 51%. For samples with a Plasmodium falciparum qPCR titer of >= 2 parasites/mu L, LAMP sensitivity was 97.8% (95% confidence interval, 93.7%-99.5%). Most false-negative LAMP results involved samples with parasitemia levels detectable by 3-well nested PCR but very low or undetectable by qPCR. Conclusions. Malaria LAMP in a remote Ugandan clinic achieved sensitivity similar to that of single-well nested PCR in a United Kingdom reference laboratory. LAMP dramatically lowers the detection threshold achievable in malaria-endemic settings, providing a new tool for diagnosis, surveillance, and screening in elimination strategies.
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