Simple detection of single nucleotide polymorphism in Plasmodium falciparum by SNP-LAMP assay combined with lateral flow dipstick.
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 N51I mutation and antifolate resistance. The PfSNP-LAMP method was validated with genomic DNA samples and parasite lysates prepared from sensitive and pyrimethamine resistant strains of P. falciparum.
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Suganya
S
Yongkiettrakul
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Jantana
J
Kampeera
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Wanwisa
W
Chareanchim
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Roonglawan
R
Rattanajak
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Wichai
W
Pornthanakasem
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand.
Wansika
W
Kiatpathomchai
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, 113 Phahonyothin Road, Klong Nueng sub-district, Klong Luang district, Pathum Thani 12120, Thailand. Electronic address: darin@biotec.or.th.
Darin
D
Kongkasuriyachai
D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D005492:Folic Acid; D018503:Genome, Protozoan; D005838:Genotype; D016778:Malaria, Falciparum / Q000175:diagnosis; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011739:Pyrimethamine / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D013048:Specimen Handling; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D013940:Thymidylate Synthase / Q000235:genetics
D017931:DNA Primers; D016054:DNA, Protozoan; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase; D011739:Pyrimethamine
Dihydrofolate reductase; Malaria detection; Plasmodium falciparum; Pyrimethamine resistant; SNP-LAMP; Single nucleotide polymorphism-loop mediated isothermal amplification