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Author Passler, S.; Pfeffer, M.
Title Detection of antibodies to alphaviruses and discrimination between antibodies to eastern and western equine encephalitis viruses in rabbit sera using a recombinant antigen and virus-specific monoclonal antibodies Type Journal Article
Year 2003 Publication Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health Abbreviated Journal J Vet Med B Infect Dis Vet Public Health
Volume (down) 50 Issue 6 Pages 265-269
Keywords Animals; Antibodies, Monoclonal/*immunology; Antibodies, Viral/*analysis/blood; DNA Primers; Encephalitis Virus, Eastern Equine/genetics/*immunology; Encephalitis Virus, Western Equine/genetics/*immunology; Encephalomyelitis, Equine/*diagnosis/*virology; Epitopes; Fluorescent Antibody Technique/*veterinary; Horses; Rabbits; Recombination, Genetic; Reverse Transcriptase Polymerase Chain Reaction/veterinary
Abstract Three arthropod-borne alphaviruses, western equine encephalitis viruses (WEEV), eastern equine encephalitis viruses (EEEV) and Venezuelan equine encephalitis viruses are the aetiological agents of a sometimes severe encephalomyelitis in equines and humans in the New World. With regard to the different ecology and epidemiology of these viruses, a method applied in serological screening should be able to distinguish between them as well as other related members of the genus Alphavirus in the American continent. However, this has been hampered in the past by (a) the close antigenic relationship between alphaviruses in traditional serological assays, especially in the routinely used haemagglutination-inhibition, and (b) the need of biosafety level 3 facilities to grow the viral antigens. An epitope blocking assay using an EEEV glycoprotein E1-expressing recombinant Sindbis virus and virus-specific monoclonal antibodies (mAbs) binding to the E1 of EEEV (strain NJ/60) and the E1 of Sindbis virus was established using automated flow cytometry. The test was evaluated using sera of infected and vaccinated rabbits. A cut-off value of 30% inhibition for antigenic complex-specific seroconversion was found to be sufficient for the detection of the respective infection. By using three different mAbs in parallel, we were able to detect alphavirus genus-, EEEV- and WEEV-complex-specific serum antibodies. As this test is based on the inhibition of binding of virus-specific mAbs, sera of every origin other than mouse can be tested. Thus, this assay may prove useful in the serological screening of a variety of animal species during an outbreak investigation.
Address Institute for Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, Ludwig-Maximilians-University, Munich, Germany
Corporate Author Thesis
Publisher Place of Publication Editor
Language English Summary Language Original Title
Series Editor Series Title Abbreviated Series Title
Series Volume Series Issue Edition
ISSN 0931-1793 ISBN Medium
Area Expedition Conference
Notes PMID:14628996 Approved no
Call Number Equine Behaviour @ team @ Serial 2639
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