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Antimicrobial Agents and Chemotherapy, July 2004, p. 2581-2587, Vol. 48, No. 7
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.7.2581-2587.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Real-Time PCR for Dihydrofolate Reductase Gene Single-Nucleotide Polymorphisms in Plasmodium vivax Isolates

E.A.3732, Parasitologie, Mycologie Médicale et Pathologie Exotique, Faculté de Médecine, Université Claude Bernard,¹ Service de Parasitologie, Mycologie Médicale et Maladies Tropicales, Hôpital E. Herriot, Hospices Civils de Lyon,² Service de Parasitologie et Maladies Tropicales, Hôpital de la Croix-Rousse, Lyon, France,⁷ Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy,³ Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, Thailand,⁴ Department of Parasitology, Faculty of Medicine, University of Indonesia, Jakarta,⁵ Deutsche Gesellschaft für Technische Zusammenarbeit (GTZ), GTZ Sis-Kes Kupang, NTT, Indonesia⁶

Received 23 January 2004/ Returned for modification 13 February 2004/ Accepted 5 March 2004

Mutations in the dhfr gene of Plasmodium vivax (pvdhfr) are associated with resistance to the antifolate antimalarial drugs. Polymorphisms in the pvdhfr gene were assessed by hybridization probe technology on the LightCycler instrument with 134 P. 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 the pvdhfr genes were found in all Thai samples (100%). pvdhfr mutant-type alleles were significantly more common in samples from travelers (42%) than in those from patients from Indonesia (5%). Surprisingly, the pvdhfr single-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 of P. vivax malaria by the World Health Organization and the Malaria National Programs.

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