Real-Time PCR for Dihydrofolate Reductase Gene Single-Nucleotide Polymorphisms inPlasmodium vivaxIsolates

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Abstract
Mutations in thedhfrgene ofPlasmodium vivax(pvdhfr) are associated with resistance to the antifolate antimalarial drugs. Polymorphisms in thepvdhfrgene were assessed by hybridization probe technology on the LightCycler instrument with 134P. vivax-infected blood samples from Turkey (n= 24), Azerbaijan (n= 39), Thailand (n= 16), Indonesia (n= 53), and travelers (n= 19). Double mutations (S58R and S117N) or quadruple mutations (F57L/I, S58R, T61M, and S117N) in thepvdhfrgenes were found in all Thai samples (100%).pvdhfrmutant-type alleles were significantly more common in samples from travelers (42%) than in those from patients from Indonesia (5%). Surprisingly, thepvdhfrsingle-mutation allele (S117N) was identified at a high frequency in parasites from Turkey and Azerbaijan (71 and 36%, respectively), where sulfadoxine-pyrimethamine is not recommended for the treatment ofP. vivaxmalaria by the World Health Organization and the Malaria National Programs.