|Hardegger, R - Prionics Ag|
|Purro, M - Prionics Ag|
|Schroder, B - Prionics Ag|
|Moyen, J - Department Of Agriculture Analysis Laboratory|
|Gormley, E - University College - Ireland|
|Vordermeier, H - Veterinary Laboratories Agency (VLA)|
|Raeber, A - Prionics Ag|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 10/7/2009
Publication Date: 10/7/2009
Citation: Hardegger, R., Purro, M., Schroder, B., Moyen, J.L., Gormley, E., Waters, W.R., Palmer, M.V., Thacker, T.C., Vordermeier, H.M., Raeber, A.J. 2009. Evaluation of a Second Generation BOVIGAM Interferon Gamma (IFN-gamma) Assay with Alternative Antigens for Stimulation of Whole Blood Cultures [abstract].
Technical Abstract: BOVIGAM®, a rapid laboratory assay, measures gamma interferon (IFN-gamma) production in whole blood samples after induction of a cell-mediated immune response (CMI) with M. bovis antigens. The test is widely used in the field and its excellent performance in TB eradication programs in many countries worldwide is well documented. Traditional use of Bovigam is based on measuring the difference in IFN-gamma production between stimulation with bovine tuberculin and avian tuberculin. Recent advances in the use of Bovigam include the application of alternative antigens for stimulation. In this setting, absolute levels of IFN-gamma are measured after stimulation with an antigen cocktail. We found that for the incorporation of the use with alternative antigens, the test can benefit from a higher range of the IFN-gamma detection part of the assay. In this study, we present several improvements of the BOVIGAM resulting in a lower detection limit for IFN-gamma, more flexibility with regard to incorporation of different reagents for stimulation (PPDs, alternative antigens) and improved ease-of-use. Inter- and intraplate variances could be reduced to < 5%. In combination with alternative antigen cocktails to be used for stimulation of the cell mediated immune response, a significant higher specificity and equivalent sensitivity in comparison with PPDs could be achieved. Furthermore, the second generation BOVIGAM assay uses a one component substrate and less washing steps and thus reduces time and cost and allows full automation for high throughput testing needs. In conclusion, the new IFN-gamma assay can be used as in improved tool for the detection of TB infected cattle.