Rapid genotyping of loci involved in antifolate drug resistance in Plasmodium falciparum by primer extension

Shalini Nair, Alan Brockman, Lucy Paiphun, François Nosten, Tim J.C. Anderson

    Research output: Contribution to journalArticle

    38 Scopus citations

    Abstract

    Current methods used to genotype point mutations in Plasmodium falciparum genes involved in resistance to antifolate drugs include restriction digestion of PCR products, allele-specific amplification or sequencing. Here we demonstrate that known point mutations in dihydrofolate reductase and dihydropteroate synthase can be scored quickly and accurately by single-nucleotide primer extension and detection of florescent products on a capillary sequencer. We use this method to genotype parasites in natural infections from the Thai-Myanmar border. This approach could greatly simplify large-scale screening of resistance mutations of the type required for evaluating and updating antimalarial drug treatment policies. The method can be easily adapted to other P. falciparum genes and will greatly simplify scoring of point mutations in this and other parasitic organisms.

    Original languageEnglish (US)
    Pages (from-to)852-858
    Number of pages7
    JournalInternational Journal for Parasitology
    Volume32
    Issue number7
    DOIs
    StatePublished - Jun 20 2002

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    Keywords

    • Dihydrofolate reductase
    • Dihydropteroate synthase
    • Plasmodium falciparum
    • Primer extension
    • Thailand

    ASJC Scopus subject areas

    • Parasitology
    • Infectious Diseases

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