High Degree of Plasmodium vivax Diversity in the Peruvian Amazon Demonstrated by Tandem Repeat Polymorphism Analysis

Margaret Kosek Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Pablo P. Yori Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Robert H. Gilman Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Maritza Calderon Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Mirko Zimic Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Raul Chuquiyauri Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Cesar Jeri Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Viviana Pinedo-Cancino Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Michael A. Matthias Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Alejandro Llanos-Cuentas Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Joseph M. Vinetz Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Laboratory of Infectious Diseases and Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benéfica PRISMA, Lima, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

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Molecular tools to distinguish strains of Plasmodium vivax are important for studying the epidemiology of malaria transmission. Two sets of markers—tandem repeat (TR) polymorphisms and MSP3α—were used to study Plasmodium vivax in patients in the Peruvian Amazon region of Iquitos. Of 110 patients, 90 distinct haplotypes were distinguished using 9 TR markers. An MSP3α polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using HhaI and AluI revealed 8 and 9 profiles, respectively, and 36 profiles when analyzed in combination. Combining TR and PCR-RFLP markers, 101 distinct molecular profiles were distinguished among these 110 patients. Nine TR markers arrayed along a 100 kB stretch of a P. vivax chromosome containing the gene for circumsporozoite protein showed non-linear linkage disequilibrium (ISA = 0.03, P = 0.001). These findings demonstrate the potential use of TR markers for molecular epidemiology studies.

Author Notes

*Address correspondence to Joseph M. Vinetz, University of California, San Diego, Division of Infectious Diseases, Department of Medicine, 9500 Gilman Drive, Mail Code 0741, George Palade Building, Cellular and Molecular Medicine West, Room 125, La Jolla, CA 92093-0741. E-mail: jvinetz@ucsd.edu

Financial Support: This work was supported by grants from the United States Public Health Service, National Institutes of Health K24AI06803, D43 TW007120, R01067727, 1U19AI089681 (JMV), and K01TW005717 (MK), an Innovations in Clinical Research Award from the Doris Duke Charitable Foundation (JMV), and a pilot project grant from the Johns Hopkins Malaria Institute (RHG).

Authors' addresses: Margaret Kosek, Pablo P. Yori, and Robert H. Gilman, Johns Hopkins Bloomberg School of Public Health, Department of International Health, Baltimore, MD, E-mails: mkosek@jhsph.edu, pyori@jhsph.edu, and gilmanbob@gmail.com. Maritza Calderon, AB Prisma, Lima, Peru, E-mail: mmcalderons@yahoo.es. Mirko Zimic, Raul Chuquiyauri, Cesar Jeri, and Viviana Pinedo-Cancino, Alejandro Llanos-Cuentas, Universidad Cayetano Heredia, Lima, Peru, E-mails: mirko.zimic@upch.edu, raulharo@ucsd.edu, cesajeri@yahoo.es, vivi_cancino@yahoo.com, and elmer.llanos@upch.edu. Michael A. Matthias and Joseph M. Vinetz, University of California San Diego, Department of Medicine, La Jolla, CA, E-mails: mmatthias@ucsd.edu and jvinetz@ucsd.edu.

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