|Adams, D. scott|
|Knowles, Donald - Don|
Submitted to: Clinical and Diagnostic Laboratory Immunology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 1/10/2003
Publication Date: 3/20/2003
Citation: Herrmann, L.M., Cheevers, W.P., McGuire, T.C., Adams, D.S., Hutton, M.M., Gavin W.G., Knowles D.P. Competitive-inhibition enzyme-linked immunosorbent assay for detection of serum antibodies to caprine arthritis-encephalitis virus: Diagnostic tool for successful eradication. Clinical and Diagnostic Laboratory Immunology. 2003. v. 10(2). p. 267-271. Interpretive Summary: Caprine Arthritis-Encephalitis Virus (CAEV) is a lentivirus capable of causing severe arthritis and mastitis in some goats. Therefore, improved diagnostic tests for the detection of CAEV infection are necessary. This study describes the validation of a competitive enzyme linked immunosorbant assay (cELISA) for detection of serum antibodies to CAEV. The high sensitivity (100%) and specificity (96.4%) of this new test makes this test an extremely useful tool for successful eradication of CAEV.
Technical Abstract: A competitive-inhibition enzyme linked immunosorbent assay (cELISA) was evaluated for detection of serum antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) in goats. This assay utilized 96 well microtiter plates containing CAEV-63 SU captured by monoclonal antibody (mAb) F7-299 and measures competitive displacement of horseradish peroxidase-conjugated mAb GPB 74A binding by undiluted goat sera (Özyörük, F., W.P. Cheevers, G.A. Hullinger, T.C. McGuire, M. Hutton, and D.P. Knowles. 2001. Clin. Diag. Lab. Immunol. 8:44-51). Two hundred sera from goats in the United States were used to determine the sensitivity and specificity of cELISA based on immunoprecipitation (IP) of [35S] methionine-labeled viral antigens as a standard of comparison. A positive cELISA test was defined as > 33.2 percent inhibition (% I) of mAb 74A binding based on two standard deviations above the mean % I of 140 IP-negative sera. At this cutoff value, there were 0/60 false-negative sera (100% sensitivity) and 5/140 false-positive sera (96.4% specificity). Additional studies utilized IP-monitored cELISA to establish a CAEV-free herd of 1640 dairy goats.