4.2 Article

Simple detection of single nucleotide polymorphism in Plasmodium falciparum by SNP-LAMP assay combined with lateral flow dipstick

Journal

PARASITOLOGY INTERNATIONAL
Volume 66, Issue 1, Pages 964-971

Publisher

ELSEVIER IRELAND LTD
DOI: 10.1016/j.parint.2016.10.024

Keywords

Plasmodium falciparum; Malaria detection; Pyrimethamine resistant; Dihydrofolate reductase; Single nucleotide polymorphism-loop mediated isothermal amplification; SNP-LAMP

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Funding

  1. National Center for Genetic Engineering and Biotechnology [P-11-00646]
  2. National Center for Science and Development Agency, Pathum Thani, Thailand

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The significant strides made in reducing global malaria burden over the past decades are being threatened by the emergence of multi-drug resistant malaria. Mechanisms of resistance to several classes of antimalarial drugs have been linked to key mutations in the Plasmodium falciparum genes. Pyrimethamine targets the dihydrofolate reductase of the bifunctional dihydrofolate reductase thymidylate synthase (DHFR-TS), and specific point mutations in the dhfr-ts gene have been assigned to resistant phenotypes. Several molecular methods are available to detect the mutant genotypes including DNA sequencing and PCR-based methods. In this study, we report the development of PfSNP-LAMP to detect nucleotide polymorphism in the dhfr gene associated with N511 mutation and antifolate resistance. The P./SNP-LAMP method was validated with genomic DNA samples and parasite lysates prepared from sensitive and pyrimethamine resistant strains of P. falciparum. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

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