2016 ACVIM Forum Research Abstract Program

The FeLV microtiter plate format ELISA method validated in this study facilitates high throughput operation by combining parallel fluid transfer processes with automated data acquisition and results interpretation. The assay method is based on sequential orthogonal screening and confirmatory protocols. The screening protocol utilizes two distinct anti-FeLV p27 mouse monoclonal antibodies that produce a colorimetric response in samples containing FeLV p27 antigen. The confirmatory protocol requires neutralization of positive samples in a separate set of controlled assays using an anti-FeLV polyclonal antibody that blocks binding of the mouse monoclonal antibodies to FeLV p27 antigen and thereby inhibits color generation. The confirmatory step offers increased assurance of specificity by enabling discrimination between infected true positives and false positives associated with several factors including anti-mouse heterophilic antibodies in patient samples.