Application of loop-mediated isothermal amplification assay combined with lateral flow dipstick for detection of Plasmodium falciparum and Plasmodium vivax

作者：Yongkiettrakul Suganya; Jaroenram Wansadaj; Arunrut Narong; Chareanchim Wanwisa; Pannengpetch Supicha; Suebsing Rungkarn; Kiatpathomchai Wansika; Pornthanakasem Wichai; Yuthavong Yongyuth; Kongkasuriyachai Darin^*

来源：Parasitology International, 2014, 63(6): 777-784.

DOI：10.1016/j.parint.2014.06.004

• Loop-mediated isothermal amplification
• Lateral flow dipstick
• Plasmodium falciparum
• Plasmodium vivax

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.

• 出版日期2014-12