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Am. J. Trop. Med. Hyg., 80(5), 2009, pp. 734-736
Copyright © 2009 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Novel dhps and pfcrt Polymorphisms in Plasmodium falciparum Detected by Heteroduplex Tracking Assay

Jonathan J. Juliano*, David J. Bacon, Jianbing Mu, Xinhua Wang, AND Steven R. Meshnick
Division of Infectious Diseases, University of North Carolina, School of Medicine, Chapel Hill, North Carolina; US Naval Medical Research Center Detachment, Lima, Peru; Laboratory of Malaria and Vector Research, National Institutes of Health, Rockville, Maryland; Guangzhou University of Traditional Chinese Medicine, Guangzhou, China; Department of Epidemiology, University of North Carolina, Gillings School of Global Public Health, Chapel Hill, North Carolina

 

ABSTRACT

Several single nucleotide polymorphisms (SNPs) have been linked to antimalarial drug resistance in Plasmodium falciparum. However, standard polymerase chain reaction (PCR) methods to detect these polymorphisms are unable to detect SNPs in variants representing < 20% of the parasites in a mixed infection, nor can they detect polymorphisms at nearby loci. Here we use heteroduplex tracking assays (HTAs) to analyze dhps540 mutations in 96 samples from Peru and pfcrt76 mutations in 70 samples from China. All samples had been previously analyzed by standard PCR. We detected drug-resistant minority variants and two novel non-synonymous pfcrt mutations in China. In Peru, we found no drug-resistant minority variants and a synonymous mutation in dhps. Thus, even in regions of low malaria transmission, HTA assays are more informative than PCR with agarose gel electrophoresis.

Received October 28, 2008. Accepted for publication January 21, 2009.

Acknowledgments: The authors thank Thomas E. Wellems for providing advice and assistance on the manuscript. One of the authors is a military service member (DJB). This work was prepared as part of his official duties.

Financial support: This project was funded in part by NIH 1R21AI07-6785 and the 2007 IDSA ERF/NFID Merle A. Sande/Pfizer Fellowship in International Infectious Diseases, KL2RR025746 from the National Center for Research Resources, and funds provided by the Department of Defense–Global Emerging Infectious System (DoD-GEIS) under Unit 847705.82000.25GB.B0016. 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.

Disclaimer: The views expressed in this article are those of the authors and do not necessarily reflect the official policy of the Department of the Navy, Department of Defense, nor the US government. Title 17 U.S.C. § 105 provides that "Copyright protection under this title is not available for any work of the US Government." Title 17 U.S.C. § 101 defines a US Government work as a work prepared by a military service member or employee of the US Government as part of that person’s official duties. The study was conducted following a protocol approved by the Walter Reed Army Institute of Research Human Use and Review Committee (Protocol 719).

* 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 J. 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. David J. Bacon, Navy Environmental and Preventative Medicine Unit-2, 1887 Powhatan St., Norfolk, VA 23511-3394, Tel: 757-953-6571, Fax: 757-953-7212, E-mail: david. bacon{at}med.navy.mil. Jianbing Mu, Laboratory of Malaria and Vector Research, National Institutes of Health, 12735 Twinbrook Parkway, MSC 8132, Twinbrook III, Room 3E-10A, Bethesda, MD 20892-8132, Tel: 301-402-1274, Fax: 301-402-2201, E-mail: jmu{at}niaid.nih.gov. Xinhua Wang, Guangzhou University of Traditional Chinese Medicine, University City, Panyu District, Guangzhou 510006, People’s Republic of China, Tel: 86-20-39358328, Fax: 86-20-39359999, E-mail: xinhuaw{at}gzhtcm.edu.cn. 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-mail: meshnick{at}email.unc.edu.






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