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Am. J. Trop. Med. Hyg., 78(6), 2008, pp. 892-894
Copyright © 2008 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Detection of the Dihydrofolate Reductase–164L Mutation in Plasmodium falciparum Infections from Malawi by Heteroduplex Tracking Assay

Jonathan J. Juliano*, Paul Trottman, Victor Mwapasa, AND Steven R. Meshnick
Division of Infectious Diseases, University of North Carolina, School of Medicine, Chapel Hill, North Carolina; Department of Epidemiology, University of North Carolina, School of Public Health, Chapel Hill, North Carolina; Department of Community Health, College of Medicine, University of Malawi, Blantyre, Malawi

 

ABSTRACT

Standard polymerase chain reaction methods often cannot detect drug-resistance mutations in Plasmodium falciparum infections if the mutation is present in ≤ 20% of the parasites. A heteroduplex tracking assay was developed that can detect dihydrofolate reductase 164-L mutations in variants representing 1% of the parasites in an individual host. Using this assay, we confirmed the presence of the mutation in P. falciparum infections in Malawi.

Received December 13, 2007. Accepted for publication March 17, 2008.

Acknowledgments: We are grateful to Ella Nkhoma and Alisa Alker for performing the real-time PCR on clinical samples.

Financial support: This study was supported by ASPH/CDC S1935-21/21, NIAID/NIH AI49804, Infectious Disease Pathogenesis Research Training grant (DHHS/NIH/NIAID 5 T32 AI07151-29), and the 2007 IDSA ERF/NFID Merle A. Sande/Pfizer Fellowship in International Infectious Diseases. These agencies had no involvement in the design, collection, analysis, or interpretation of data in this study or in writing this paper or submitting it for publication.

* Address correspondence to Jonathan J. Juliano, Division of Infectious Diseases, University of North Carolina, School of Medicine, CB#7300, Chapel Hill, NC 27599-7300. E-mail: jjuliano{at}med.unc.edu

Authors’ addresses: Jonathan Juliano, Division of Infectious Diseases, University of North Carolina, School of Medicine, CB#7300, Chapel Hill, NC 27599-7300, Tel: 919-843-4384, Fax: 919-966-0584, E-mail: jjuliano{at}med.unc.edu. Paul Trottman and Steven R. Meshnick, Department of Epidemiology, University of North Carolina, School of Public Health, CB#7435, Chapel Hill, NC 27599-7435, Tel: 919-843-4384, Fax: 919-966-0584, E-mails: ptrottman{at}unc.edu and meshnick{at}email.unc.edu. Victor Mwapasa, College of Medicine, University of Malawi, Mahatma Gandhi Campus, Private Bag 360, Chichiri, Blantyre 3, Malawi, Tel: 265-8-206-255, E-mail: vmwapasa{at}medcol.mw.




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Plasmodium falciparum Strains Harboring Dihydrofolate Reductase with the I164L Mutation Are Absent in Malawi and Zambia Even under Antifolate Drug Pressure
Antimicrob. Agents Chemother., November 1, 2008; 52(11): 3883 - 3888.
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Copyright © 2008 by the American Society of Tropical Medicine and Hygiene.