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Am. J. Trop. Med. Hyg., 74(4), 2006, pp. 546-553
Copyright © 2006 by The American Society of Tropical Medicine and Hygiene

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POLYMERASE CHAIN REACTION AND MOLECULAR GENOTYPING TO MONITOR PARASITOLOGICAL RESPONSE TO ANTI-MALARIAL CHEMOTHERAPY IN THE PERUVIAN AMAZON

EVERICK AYALA{dagger}, ANDRÉS G. LESCANO{dagger}, ROBERT H. GILMAN, MARITZA CALDERÓN, VIVIANA V. PINEDO, HILJA TERRY, LILIA CABRERA, AND JOSEPH M. VINETZ*
Asociación Benefica PRISMA, Lima, Peru; Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Infectious Diseases Laboratory, Department of Pathology, Universidad Peruana Cayetano Heredia, Lima, Peru; Direccion de Salud, Ministry of Health, Loreto Department, Iquitos, Peru; Division of Infectious Diseases, Department of Medicine, University of California, San Diego School of Medicine, La Jolla, California

Over the past decade, anti-malarial drug resistance has rapidly become a major public health problem in the Peruvian Amazon. This study compared polymerase chain reaction (PCR) to light microscopy for diagnosing and monitoring the parasitological response of malaria patients to anti-malarial chemotherapy in the Peruvian Amazon region of Iquitos. Typing of P. falciparum using MSP1, MSP2, and glutamine-rich protein distinguished among infecting parasites. Most (73%) P. falciparum patients were parasitologically resistant to sulfadoxine-pyrimethamine (RI = 10, RII = 1). Sensitivity of microscopy was lower than PCR (69% for P. vivax and 78% for P. falciparum), but parasite clearance times were comparable between microscopy and PCR. PCR sensitively and specifically detected mixed infections and low-level parasitemia indicative of drug resistance, making this approach of practical use for the control of malaria at the public health level. Genotyping malaria parasites will be useful to distinguish drug failure from new infections in clinical trials of anti-malarial drugs in the Peruvian Amazon region.

Received December 6, 2004. Accepted for publication January 6, 2006.

Acknowledgments: We thank the patients and staff of the health post of Varillal, especially César Banda for his invaluable assistance in microscopy readings; Dr. Carlos Calampa, Chief of Loreto Health Directorate, for his cooperation; Guillermo Achón, Loreto Health Directorate, for his contributions in the field of work; and Consuelo Ludeña, Pola Valles, Michael Levy, J. B. Phu, and D. Sara for their technical assistance.

Disclaimer: The views expressed in this article are those of the contributing author Andrés G. Lescano and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the U.S. Government.

Financial support: This study was funded in part by USAID HRN-A-00-96-90006-00 GRANT, by an ITREID Fogarty training grant, a Doris Duke Charitable Foundation Innovations in Clinical Research Award, and the anonymous RG-ER fund.

* Address correspondence to Joseph M. Vinetz, University of California San Diego, School of Medicine, 9500 Gilman Drive, 0640 La Jolla, CA 92093-0640. E-mail: jvinetz{at}ucsd.edu

{dagger} These authors contributed equally to this study.

Authors’ addresses: Everick Ayala, Robert H. Gilman, Hilja Terry, Lilia Cabrera, Asociación Benefica PRISMA, Carlos Gonzales 251 Urb. Maranga San Miguel, Lima 32, Peru, E-mail: everickayla{at}yahoo.com, rgilman{at}jhsph.edu, hilaterry{at}yahoo.com, and lcabrera{at}peruresearch.com. Andrés G. Lescano, United States Navy Medical Research Center Detatchment, Peru, NMRCD Unit 3800, American Embassy–Lima, APO AA 34031, Telephone: 511-562-3848 ext 144, Fax: 511-562-3833, E-mail: wlescano{at}hotmail.com. Andrés G. Lescano, Robert H. Gilman, Bloomberg School of Public Health, Department of International Health, The Johns Hopkins University, 615 N. Wolfe Street, Room W3501 Baltimore, MD 21205-2179. Maritza Calderón, Viviana V. Pinedo, Infectious Diseases Laboratory, Department of Pathology, Universidad Peruana Cayetano Heredia. Av. Honorio Delgado 430, Urb. Ingeniería, Lima 31, E-mail: mmcalderons{at}yahoo.com and viviana_cancino{at}yahoo.com. Joseph M. Vinetz, Division of Infectious Diseases, Department of Medicine, University of California San Diego School of Medicine, Center for Molecular Medicine East-Room 2052, 9500 Gilman Drive, La Jolla, CA 92093-0640. Alexander von Humboldt Institute of Tropical Medicine, Universidad Peruana Cayetano Heredia, Av. Honorio Delgado 430, Urb. Ingeniería, Lima 31, Peru, E-mail: jvinetz{at}ucsd.edu.




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