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Technical Briefs |
1 Department of Microbiology, The University of Hong Kong, Hong Kong SAR;2 Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong SAR;3 Department of Parasitology and Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Sri Lanka;4 Department of Clinical Tropical medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand;
aaddress correspondence to this author at: Department of Microbiology, University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong SAR; fax 852-2855-1241, e-mail llmpoon{at}hkucc.hku.hk
Abstract
Background: Malaria is one of the most important parasitic infections in humans. A sensitive diagnostic test for malaria that could be applied at the community level could be useful in programs to control the disease. The aim of the present work was to develop a simple, inexpensive molecular test for Plasmodium falciparum.
Methods: Blood was collected from controls (n = 100) and from patients diagnosed with falciparum malaria infection (n = 102), who were recruited to the study. Heat-treated blood samples were tested by a loop-mediated isothermal amplification (LAMP) assay for P. falciparum. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. To evaluate the assay, DNA from these samples was purified and tested by PCR. Results from the LAMP and PCR assays were compared.
Results: The LAMP assay detected P. falciparum directly from heat-treated blood. The quantitative data from the assay correlated to the parasite counts obtained by blood-film microscopic analyses. When we used the PCR assay as the comparison method, the sensitivity and specificity of the LAMP assay were 95% and 99%, respectively.
Conclusions: Unlike PCR, the LAMP assay does not require purified DNA for efficient DNA amplification, thereby reducing the cost and turnaround time for P. falciparum diagnosis. The assay requires only basic instruments, and assay positivity can be verified by visual inspection.
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