1921
Volume 76, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

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.

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2007-03-01
2017-11-20
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References

  1. Meegan JM, Digoutte JP, Peters CJ, Shope RE, 1983. Monoclonal antibodies to identify Zinga virus as Rift Valley fever virus. Lancet 1 : 641.
  2. Turell MJ, O’Guinn ML, Jones JW, Sardelis MR, Dohm DJ, Watts DM, Fernandez R, Travassos Da Rosa A, Guzman H, Tesh R, Rossi CA, Ludwig GV, Mangiafico JA, Kondig J, Wasieloski LP Jr, Pecor J, Zyzak M, Schoeler G, Mores CN, Calampa C, Lee JS, Klein TA, 2005. Isolation of viruses from mosquitoes collected in the Amazon Basin region of Peru. J Med Entomol 42 : 891–898.
  3. Morris CD, 1989. Eastern equine encephalitis. Monath TP, ed. The Arboviruses: Epidemiology and Ecology. Volume III. Boca Raton, FL: CRC Press, 1–20.
  4. Weaver SC, Hagenbaugh A, Bellew LA, Gousset L, Mallampalli V, Holland JJ, Scott TW, 1994. Evolution of alphaviruses in the eastern equine encephalomyelitis complex. J Virol 68 : 158–169.
  5. Brault AC, Powers AM, Chavez CL, Lopez RN, Cachon MF, Gutierrez LF, Kang W, Tesh RB, Shope RE, Weaver SC, 1999. Genetic and antigenic diversity among eastern equine encephalitis viruses from North, Central, and South America. Am J Trop Med Hyg 61 : 579–586.
  6. Lennette EH, Schmidt NJ, eds, 1964. Diagnostic Procedures for Viral and Rickettsial Diseases: Fluorescent Antibody Techniques. New York: American Public Health Association, Inc., 177–193.
  7. Thompson JD, Higgins DG, Gibson TJ, 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 : 4673–4680.
  8. Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD, 2003. Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31 : 3497–3500.
  9. Howe K, Bateman A, Durbin R, 2002. QuickTree: building huge neighbor-joining trees of protein sequences. Bioinformatics 18 : 1546–1547.
  10. Hahn CS, Lustig S, Strauss EG, Strauss JH, 1988. Western equine encephalitis virus is a recombinant virus. Proc Natl Acad Sci USA 85 : 5997–6001.
  11. LaStarza MW, Lemm JA, Rice CM, 1994. Genetic analysis of the nsP3 region of Sindbis virus: evidence for roles in minus-strand and subgenomic RNA synthesis. J Virol 68 : 5781–5791.
  12. O’Guinn ML, Lee JS, Kondig JP, Fernandez R, Carbajal F, 2004. 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. Am J Trop Med Hyg 70 : 164–171.
  13. Kuno G, 1998. Universal diagnostic RT-PCR protocol for arboviruses. J Virol Methods 72 : 27–41.
  14. Martinez HM, 1983. An efficient method for finding repeats in molecular sequences. Nucleic Acids Res 11 : 4629–4634.
  15. Needleman SB, Wunsch CD, 1970. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol 48 : 443–453.
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  • Received : 17 Feb 2006
  • Accepted : 04 Dec 2006

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