Development and validation of two competitive ELISAs for detecting human and canine adenovirus antibody in Ontario wildlife sera

by Maxwell, Laura

A human adenovirustype 5-rabies recombinant vaccine(Ad5RG) is a potential candidateforthe oral immunization ofskunks, foxes, and raccoons against rabies. Adenovirus seroprevalence studies in these species were required before using the vaccine in the field. as part of an environmental risk assessrnentand for determiningif a barrier to immunizationexists. For this purpose, two cornpetitive enzyme-linked immunosorbentassays (CELISAS) rneasuring antibodies (Ab) against Ad5 and canine adenovims type one (CAV- 1) were developed and validated. Referencesera used forvalidation were derived from Iaboratoryskunks. foxes. and raccoons glven Ad5RG by eithertheoralor intra-muscularroute. For CAV- 1 cELISA validation. additional sera wereused from dog vaccinated intra-muscularl y with CXV-2. which protects asainst CAV- I and C AV-2 infection. Sera were examined For either Ad5 or C AV- 1 neutnlizing antibodies using plaque reduction neutralization tests (PRNT) .As part ofcELISA development, 1O Ad5 and IO CAV- i mAbs were characterized accordingto their viral protein specificities. their neutralizing abilities, and theircross-reactions withother adenoviruses. An Ad5 non-neutralizing hexon m .Ab (1 5G6) that cross-reactedwith Ad 1-30was selected forthe Ad5 CELIS