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Although genetically distinct malaria parasites have been shown to simultaneously infect an individual, the total number of unique parasites has not been systematically studied. We examined multiple clones (8–38) from individual blood samples collected from Papua New Guinean children for polymorphisms in the Plasmodium vivax Duffy binding protein (dbpII) and the merozoite surface protein 3
(msp3). We found a median of 4 (range = 2–6) and 12 (range = 2–23) unique genotypes based on dbpII and msp3, respectively, per person at one time point and at least 12–33 unique genotypes per person over a four-month period. Control polymerase chain reactions (PCRs) detected 0–31% of clones with haplotypes that arose from PCR artifacts, indicating that caution must be taken when using PCR-based analysis to examine complex infections. To reduce artifacts from clones, analysis was based on haplotypes unlikely to have been generated by PCR artifacts or had been previously identified. Plasmodium vivax infections can be highly complex in disease-endemic areas, suggesting continual genetic mixing that could have significant implications for the use of antimalarial drugs and malaria vaccines.
Received January 10, 2005. Accepted for publication May 11, 2005.
Acknowledgments: We thank the study participants for their time and P. A. Zimmerman for helpful discussions and suggestions.
Financial support: This study was supported by a grant from the Veterans Affairs Research Service.
* Address correspondence to Jennifer L. Cole-Tobian or Christopher L. King, Center for Global Health and Disease, Case Western Reserve University, Wolstein Research Building, Rm. 4132, 2103 Cornell Rd., Cleveland, OH 44106. E-mail: jlc11{at}cwru.edu, christopher.king{at}case.edu
Authors’ addresses: Jennifer L. Cole-Tobian, Center for Global Health and Disease, Case Western Reserve University, Wolstein Research Building 4-101, 10900 Euclid Avenue, Cleveland, OH 44106, E-mail: jlc11{at}cwru.edu. Moses Biasor, Papua New Guinea Institute of Medical Research, PO Box 378, Madang 511, Papua New Guinea. Christopher L. King, Center for Global Health and Disease, Case Western Reserve University, Wolstein Research Building 4132, 2103 Cornell Rd., Cleveland, OH 44106 and Research Department, Veteran’s Affairs Medical Center, 10701 East Boulevard, Cleveland, OH 44106, Telephone: 216-368-4817, Fax: 216-368-4825, E-mail: christopher.king{at}case.edu.
Reprint requests: Christopher L. King, Center for Global Health and Disease, Case Western Reserve University, Wolstein Research Building 4-132, 2103 Cornell Rd., Cleveland, OH 44106.
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