Changes

*Virus may be identified in brain tissue or cell culture using direct immunofluorescent staining.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>

*Virus may be identified in brain tissue or cell culture using direct immunofluorescent staining.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>

*EEE and WEE viral RNA in mosquitoes and equine tissues may be detected by reverse-transcription PCR.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>   

*EEE and WEE viral RNA in mosquitoes and equine tissues may be detected by reverse-transcription PCR.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>   

*ELISA can be used to detect virus in brain tissue.  An antigen-capture ELISA, developed for EEE surveillance in mosquitoes, can be used where virus isolation and PCR facilities are unavailable. <ref>Brown, T.M, Mitchell, C.J, Nasci, R.S, Smith, G.C. and Roehrig, J.T. (2001). Detection of eastern equine encephalitis virus in infected mosquitoes using a monoclonal antibody-based antigen-capture enzyme-linked immunosorbent assay. ''Am. J. Trop. Med. Hyg.'', 65, 208-213.  In: ''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>

*ELISA can be used to detect virus in brain tissue.  An antigen-capture ELISA, developed for EEE surveillance in mosquitoes, can be used where virus isolation and PCR facilities are unavailable. <ref>Brown, T.M, Mitchell, C.J, Nasci, R.S, Smith, G.C. and Roehrig, J.T. (2001). Detection of eastern equine encephalitis virus in infected mosquitoes using a monoclonal antibody-based antigen-capture enzyme-linked immunosorbent assay, ''Am. J. Trop. Med. Hyg.'', 65, 208-213.  In: ''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>

*Virus isolation is the most definitive diagnostic method for EEE or WEE.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Brain is preferred, but virus has also been isolated from the liver and spleen.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>. Samples of these tissues should be taken in duplicate, one set for virus isolation and the other placed in formalin for histopathology.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>. Viral isolation specimens should be sent frozen unless they can be received refrigerated within 48 hours of sampling.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Unless clinical signs persist for more than 5days prior to death, EEE virus is frequently isolated from equine brain tissue.  WEE virus, however, is rarely isolated from tissues of infected horses.  Newborn mice, chicken embryos and a number of cell culture systems can be used for virus isolation.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Virus may also be isolated from cerebrospinal fluid (CSF) of acutely infected horses.<ref name="duplicate">Merck & Co (2008) The Merck Veterinary Manual (Eighth Edition), Merial found at http://www.merckvetmanual.com/mvm/index.jsp?cfile=htm/bc/100900.htm&word=Equine%2cencephalitis, accessed July 2010</ref>

*Virus isolation is the most definitive diagnostic method for EEE or WEE.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Brain is preferred, but virus has also been isolated from the liver and spleen.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>. Samples of these tissues should be taken in duplicate, one set for virus isolation and the other placed in formalin for histopathology.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>. Viral isolation specimens should be sent frozen unless they can be received refrigerated within 48 hours of sampling.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Unless clinical signs persist for more than 5days prior to death, EEE virus is frequently isolated from equine brain tissue.  WEE virus, however, is rarely isolated from tissues of infected horses.  Newborn mice, chicken embryos and a number of cell culture systems can be used for virus isolation.<ref name="again">''Manual of Diagnostic Tests and Vaccines for Terrestrial Animals'' found at http://www.oie.int/eng/normes/mmanual/A_00081.htm, accessed July 2010.</ref>.  Virus may also be isolated from cerebrospinal fluid (CSF) of acutely infected horses.<ref name="duplicate">Merck & Co (2008) The Merck Veterinary Manual (Eighth Edition), Merial found at http://www.merckvetmanual.com/mvm/index.jsp?cfile=htm/bc/100900.htm&word=Equine%2cencephalitis, accessed July 2010</ref>