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Identifying viral isolates from field-collected mosquitoes can be difficult and time-consuming, particularly in regions of the world where numerous closely related viruses are co-circulating (e.g., the Amazon Basin region of Peru). The use of molecular techniques may provide rapid and efficient methods for identifying these viruses in the laboratory. Therefore, we determined the complete nucleotide sequence of two South American eastern equine encephalomyelitis viruses (EEEVs): one member from the Peru-Brazil (Lineage II) clade and one member from the Argentina-Panama (Lineage III) clade. In addition, we determined the nucleotide sequence for the nonstructural P3 protein (nsP3) and envelope 2 (E2) protein genes of 36 additional isolates of EEEV from mosquitoes captured in Peru between 1996 and 2001. The 38 isolates were evenly distributed between lineages II and III virus groupings. However, analysis of the nsP3 gene for lineage III strongly suggested that the 19 isolates from this lineage could be divided into two sub-clades, designated as lineages III and IIIA. Compared with North American EEEV (lineage I, GA97 strain), we found that the length of the nsP3 gene was shorter in the strains isolated from South America. A total of 60 nucleotides was deleted in lineage II, 69 in lineage III, and 72 in lineage IIIA. On the basis of the sequences we determined for South American EEEVs and those for other viruses detected in the same area, we developed a series of primers for characterizing these viruses.
Received February 17, 2006. Accepted for publication December 4, 2006.
Acknowledgments: We thank Mark Wooster, Faustino Carbajal, Roberto Fernandez, Karla Block, and Helvio Astete (Naval Medical Research Center and Detachment-Peru) for technical support. We also thank the Peruvian Ministry of Health for their assistance; James Pecor, Richard Wilkerson, and E. L. Peyton (Walter Reed Army Medical Institute of Research) for providing taxonomic assistance; Katheryn Kenyon for expert editorial assistance; Lorraine Farinick for providing the illustrations; George Korch for his support and helpful suggestions; and the Amazon Center for Environmental Education and Research for the use of their facilities and assistance.
Financial support: This work was supported by work unit number 62787A 870 U 8517 of the U.S. Navy and by Defense Threat Reduction Agency project G0015 04 RD B.
Disclaimer: The use of trade names or commercial products does not constitute endorsement or recommendation for use by the Department of Defense. The views of the authors do not necessarily reflect the position of the Department of Defense or the Department of the Army.
* Address correspondence to Michael J. Turell, Department of Cell Biology, Virology Division, U.S. Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702-5011. E-mail: michael.turell{at}det.amedd.army.mil
Authors’ addresses: John P. Kondig and Leonard P. Wasieloski Jr, Diagnostics Systems Division, U.S. Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702-5011, E-mail: John.Kondig{at}det.amedd.army.mil. Michael J. Turell, John S. Lee, and Monica L. O’Guinn, Virology Division, U.S. Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702-5011.
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