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FIELD DETECTION OF EASTERN EQUINE ENCEPHALITIS VIRUS IN THE AMAZON BASIN REGION OF PERU USING REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION ADAPTED FOR FIELD IDENTIFICATION OF ARTHROPOD-BORNE PATHOGENS

In support of efforts to develop rapid diagnostic assays for use in the field, reverse transcription-polymerase chain reaction (RT-PCR) assays were developed to detect arboviruses circulating in the Amazon Basin region of Peru. Previous knowledge of arthropod/pathogen relationships allowed a focused evaluation to be conducted in November 2000 that assessed the feasibility and reliability of a mobile, rapid, field-expedient RT-PCR diagnostic system aimed at detecting eastern equine encephalitis virus (EEEV) in Culex (Melanoconion) pedroi mosquitoes. Modifications were made to a commercially available mobile molecular laboratory kit and assay procedures were tailored for use under harsh environmental conditions with field-collected and field-processed mosquitoes. From CO₂ baited mosquito light traps, 3,227 Cx. (Mel.) pedroi mosquitoes were collected and sorted into 117 pools. The pools were processed and assayed in the field by RT-PCR and five of those pools were found positive for EEEV. Laboratory sequence analysis confirmed the presence of two distinct subtypes of EEEV.

Received March 23, 2003. Accepted for publication November 6, 2003.

Acknowledgments: We thank Dr. Michael Turell for inoculating select mosquitoes, Angela Hadjipanayis for plaque assaying select viruses, Michael Delgado and Dieter Schachner for rearing of the laboratory mosquitoes, and the United States Navy Medical Research Center Detachment (Lima and Iquitos, Peru) for the logistical support provided during the field evaluation.

Disclaimer: The views in this report are those of the authors and should not be considered as positions of the Department of the Army.

Authors’ addresses: Monica L. O’Guinn, Command, USACHPPM-Pac, ATTN: MCHB-AJ-TEM (MAJ O’Guinn), APO, AP 96343-5006, Telephone: 81-3117-63-4478, Fax: 81-3117-63-8597. John S. Lee, Virology Division, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, Frederick, MD 21702, Telephone: 301 619-4912, Fax: 301-619-2290, E-mail: john.lee{at}det.amedd.army.mil. John P. Kondig, Diagnostics Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, Telephone: 301-619-4781, Fax: 301-619-2290. Roberto Fernandez, United States Naval Medical Research Center Detachment, Unit 3800, APO, AA 34031, Telephone: 51-1-561-2882 extension 139, Fax: 51-1-561-304. Faustino Carbajal, 4301 Columbia Pike # 515, Arlington, VA, 22204, Telephone: 703-521-2118.

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				J. P. KONDIG, M. J. TURELL, J. S. LEE, M. L. O'GUINN, and L. P. WASIELOSKI JR GENETIC ANALYSIS OF SOUTH AMERICAN EASTERN EQUINE ENCEPHALOMYELITIS VIRUSES ISOLATED FROM MOSQUITOES COLLECTED IN THE AMAZON BASIN REGION OF PERU Am J Trop Med Hyg, March 1, 2007; 76(3): 408 - 416. [Abstract] [Full Text] [PDF]