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Detection of Neutralization-sensitive Epitopes in Antigens Displayed on Virus-Like Particle (VLP)-Based Vaccines Using a Capture Assay
Published on: February 10, 2022
Detection of antibodies against capripoxviruses using an inactivated sheeppox virus ELISA
S Babiuk1, D B Wallace, S J Smith
1National Centre for Foreign Animal Disease, Winnipeg MB, Canada. shawn.babiuk@inspection.gc.ca
Abstract:
An indirect ELISA was developed to detect antibodies specific for capripoxviruses in goat, sheep and cattle sera. Heat-inactivated Nigerian sheeppox virus was used as the ELISA antigen. Sera obtained from sheep and goats that were experimentally infected with different capripoxvirus isolates were used to develop and evaluate the sensitivity of the ELISA. Virus neutralization indexes were determined for the experimental sera in OA3.Ts cells. The specificity of the ELISA was determined using 231 sera from capripoxvirus naïve sheep and goats from Canada. In addition, the ELISA was tested for cross-reactivity to anti-orf virus antibodies using orf-reactive sera and no cross-reactivity was observed. Using experimentally generated sera obtained from animals infected with virulent sheeppox or goatpox virus isolates, the diagnostic sensitivity of the ELISA was 96% with a diagnostic specificity of 95%, where the diagnostic sensitivity of the virus neutralization assay was 96% with a diagnostic specificity of 100%. Further evaluation of this ELISA, using 276 cattle serum samples that were positive by virus neutralization assays, revealed a diagnostic sensitivity of 88% with a specificity of 97%. These results indicated that the inactivated capripoxvirus ELISA can detect capripoxvirus-specific antibodies in sheep, goats and cattle that have been infected with virulent capripoxvirus isolates. Non-virulent capripoxvirus isolates, in contrast, did not elicit positive (>or=1.5 Log10 neutralization index) antibody responses.

