Journal
PARASITOLOGY INTERNATIONAL
Volume 63, Issue 6, Pages 777-784Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.parint.2014.06.004
Keywords
Loop-mediated isothermal amplification; Lateral flow dipstick; Plasmodium falciparum; Plasmodium vivax
Categories
Funding
- National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand
- National Center for Genetic Engineering and Biotechnology (BIOTEC)
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Malaria is largely a preventable and curable disease. However, a delay or an inappropriate treatment can result in serious adverse outcomes for patient. Rapid, simple and cost-effective diagnostic tests that cab be easily adapted and rapidly scaled-up at the field or community levels are needed. In this study, accelerated detection methods for the Plasmodium falciparum (Pf) and Plasmodium vivax (Pv) dihydrofolate reductase-thymidylate synthase were developed based on the loop-mediated isothermal amplification (LAMP) method. The developed methods included the use of species-specific biotinylated primers to amplify LAMP amplicons, which were then hybridized to specific FITC-labeled DNA probes and visualized on a chromatographic lateral flow dipstick (LFD). The total LAMP-LFD assay time was approximately 1.5 h. The LAMP-LFD assays showed similar detection limit to conventional PCR assay when performed on plasmid DNA carrying the malaria dhfr-ts genes. The LAMP-LFD showed 10 folds higher detection limit than PCR when performed on genomic DNA samples from Pf and Pv parasites. The dhfr-ts LAMP-LFD assays also have the advantages of reduced assay time and easy format for interpretation of results. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
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