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We developed a novel Plasmodium falciparum genotyping strategy based on the heteroduplex tracking assay (HTA) method commonly used to genotype viruses. Because it can detect both sequence and size polymorphisms, we hypothesized that HTA is more sensitive than current methods. To test this hypothesis, we compared the ability of HTA and a nested polymerase chain reaction (PCR) to detect genetic diversity in 17 Thai samples. The HTA detected more MSP1 sequence variants in eight isolates (47%), less sequence variants in three isolates (18%), and an equal number of sequence variants in six isolates (35%), suggesting that HTA is equal to or more sensitive than the nested PCR. This study is a proof of concept that HTA is a sensitive allelic discrimination method able to determine genetic diversity in P. falciparum and warrants its use in studies of antimalarial drug efficacy.
Received May 28, 2004. Accepted for publication December 8, 2004.
Acknowledgments: We thank Paul Wilson for technical support, Alisa Alker for helpful discussions, and the referees for their thorough review of this manuscript.
Financial support: This work was supported by National Institutes of Health (NIH) grants R23 AI054590 (Steven R. Meshnick), R01-AI44667 (Ronald Swanstrom), T32 AI07151-26 (Jesse J. Kwiek), NIH Training Grant T32-AI07419 (Kimberly Ritola), and a scholarship from the Faculty of Medical Technology at Mahidol University under the Thai staff development project of the Ministry of University Affairs (Warunee Ngrenngarmlert). We also acknowledge the generous support of the University of North Carolina Center for AIDS Research (P30-AI50410).
* These authors contributed equally to this article.
Authors’ addresses: Warunee Ngrenngarmlert and Wanida Ittarat, Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand, Telephone: 66-2-419-7170, Fax: 66-2-412-4110. Jesse J. Kwiek and Steven R. Meshnick, Department of Microbiology and Immunology, CB# 7290, University of North Carolina, Chapel Hill, NC 27599, Telephone: 919-843-4384, Fax: 919-962-8103, E-mail: meshnick{at}unc.edu. Deborah D. Kamwendo, Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, MI 48109. Kimberly Ritola and Ronald Swanstrom, Center for AIDS Research, University of North Carolina, Chapel Hill, NC 27599. Chansuda Wongsrichanalai and R. Scott Miller, Department of Immunology and Medicine, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
Reprint requests: Steven R. Meshnick, Department of Epidemiology, CB# 7435, University of North Carolina, Chapel Hill, NC 27599.
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