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United States Department of Agriculture

Agricultural Research Service

Title: Ovine Progressive Pneumonia Virus Capsid Is B-Cell Immunodominant Using Western Blot Analysis: a Comparison of Sensitivity Between Western Blot Analysis and Immunoprecipitation

item Myers-Evert, Dawn - WSU
item Hoesing, Lynn

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 26, 2006
Publication Date: November 1, 2006
Citation: Myers-Evert, D.K., Hoesing, L.M. 2006. Ovine progressive pneumonia virus capsid is B-cell immunodominant using Western blot analysis: A comparison of sensitivity between Western blot analysis and immunoprecipitation. Journal of Virological Methods. 137(2):339-342.

Interpretive Summary: The sensitivity of immunoprecipitation, the comparable standard for determining OPPV infection status in serum, was tested against the sensitivity of western blot analysis. It was determined that western blot analysis was 12-fold more sensitive than immunoprecipitation. Western blot analysis also requires one-fiftieth of the time to perform and does not require radioactivity. Diagnostic laboratories can perform this test without much difficulty; therefore, this test will replace immunoprecipitation as the comparable standard in the future.

Technical Abstract: A western blot assay (WB) was developed and analyzed against the comparable standard, immunoprecipitation of 35[S] methionine/cysteine-labeled ovine progressive pneumonia virus (OPPV) proteins (IP), for its ability to detect anti-OPPV antibodies using endpoint titers. WB is 12-fold more sensitive in detecting endpoint anti-capsid (CA) antibody titers than IP, and CA is the B-cell immunodominant OPPV protein when utilizing WB. Since SU is the B-cell immunodominant OPPV protein when utilizing IP, this suggests IP and WB measure different types of antibody that are more specific for conformational and linear OPPV protein epitopes, respectively.

Last Modified: 4/21/2015
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