Critical factors affecting the diagnostic reliability of enzyme-linked immunosorbent assay formats

This paper aims to evaluate different formats of the enzyme-linked immunosorbent assays (ELISAs) for detection of virus-specific antibodies and focuses on factors that may influence the diagnostic reliability of such tests. Newly developed and well-established ELISAs for detection of infections of bovine herpesvirus 1 (BHV1), bovine respiratory syncytial virus (BRSV), classical swine fever virus (CSFV), pseudorabies virus (PRV) and bovine viral diarrhoea virus (BVDV) are used as examples. Differences between competitive and non-competitive ELISAs are described, with special reference to the influence of the antigen, the conjugated antibody and the test sample on the test results. Attention is drawn to interference, which may result in false positive or false negative test results, with special emphasis on the 'bridging' phenomenon. The use of monoclonal antibodies and discriminatory tests are briefly discussed. Diagnostic reliability is described for tests that are used in monitoring or eradication programmes, emphasising the consequences of false negative and false positive test results. Finally, reducing assay-lime and functional quality control for such tests are discussed.