{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T18:27:47Z","timestamp":1771612067643,"version":"3.50.1"},"reference-count":15,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2009,8,1]],"date-time":"2009-08-01T00:00:00Z","timestamp":1249084800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Vaccine Immunol"],"published-print":{"date-parts":[[2009,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Antigens of\n            <jats:italic>Mycobacterium bovis<\/jats:italic>\n            elicit a cell-mediated immune response upon intradermal injection in cattle. In vitro, such antigens stimulate the production of gamma interferon (IFN-\u03b3) by bovine T cells in whole-blood culture (IFN-\u03b3 assay). We have analyzed various parameters of the in vitro IFN-\u03b3 assay, ranging from blood sampling to execution of the IFN-\u03b3 test, in view of potential simplifications of the assay. Here, we show that IFN-\u03b3 responses may be reduced under certain animal handling\/holding conditions and that a delayed time from blood collection to culture may lead to a reduced in vitro IFN-\u03b3 response. Delayed initiation of culture in a purified-protein-derivative-based assay (24 h compared to 8 h after blood collection), however, resulted in a significant improvement of specificity (97% compared to 85%), whereas there was only a modest reduction of sensitivity (from 96% to 90%), which was statistically not significant. Furthermore, we show that the stimulation temperature needs to be 33\u00b0C or higher; that carbon dioxide is not required for stimulation; and that various plate formats, ranging from 24 to 96 wells per plate, can be utilized. The produced IFN-\u03b3 is stable at 4\u00b0C for 28 days as well as after repeated freeze-thaw cycles. Thus, stimulation of samples may be initiated in the field without the need for a carbon dioxide source, and bovine IFN-\u03b3 is stable under various routine laboratory temperature scenarios. These findings demonstrate opportunities for improvements in the bovine IFN-\u03b3 test platform and flexibilities in test application.\n          <\/jats:p>","DOI":"10.1128\/cvi.00150-09","type":"journal-article","created":{"date-parts":[[2009,7,2]],"date-time":"2009-07-02T01:46:33Z","timestamp":1246499193000},"page":"1196-1202","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":47,"title":["Optimization of a Whole-Blood Gamma Interferon Assay for Detection of\n            <i>Mycobacterium bovis<\/i>\n            -Infected Cattle"],"prefix":"10.1128","volume":"16","author":[{"given":"Irene","family":"Schiller","sequence":"first","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W. Ray","family":"Waters","sequence":"additional","affiliation":[{"name":"National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H. Martin","family":"Vordermeier","sequence":"additional","affiliation":[{"name":"Veterinary Laboratory Agency, Addlestone, Great Britain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian","family":"Nonnecke","sequence":"additional","affiliation":[{"name":"National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Welsh","sequence":"additional","affiliation":[{"name":"AFBI, Veterinary Sciences Division, Stormont, Great Britain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicolas","family":"Keck","sequence":"additional","affiliation":[{"name":"Laboratoire De\u0301partemental Ve\u0301te\u0301rinaire de l'He\u0301rault, Montpellier, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adam","family":"Whelan","sequence":"additional","affiliation":[{"name":"Veterinary Laboratory Agency, Addlestone, Great Britain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"Sigafoose","sequence":"additional","affiliation":[{"name":"National Veterinary Services Laboratories, APHIS, U.S. Department of Agriculture, Ames, Iowa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christoph","family":"Stamm","sequence":"additional","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mitchell","family":"Palmer","sequence":"additional","affiliation":[{"name":"National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tyler","family":"Thacker","sequence":"additional","affiliation":[{"name":"National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roland","family":"Hardegger","sequence":"additional","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beatrice","family":"Marg-Haufe","sequence":"additional","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex","family":"Raeber","sequence":"additional","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruno","family":"Oesch","sequence":"additional","affiliation":[{"name":"Prionics AG, Schlieren, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01184-06"},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1136\/vr.153.20.615","volume":"153","year":"2003","unstructured":"Buddle, B. M., A. R. McCarthy, T. J. Ryan, J. M. Pollock, H. M. Vordermeier, R. G. Hewinson, P. Andersen, and G. W. de Lisle. 2003. Use of mycobacterial peptides and recombinant proteins for the diagnosis of bovine tuberculosis in skin test-positive cattle. Vet. Rec.153:615-620.","journal-title":"Vet. Rec."},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1136\/vr.160.19.660","volume":"160","year":"2007","unstructured":"Coad, M., R. G. Hewinson, D. Clifford, H. M. Vordermeier, and A. O. Whelan. 2007. Influence of skin testing and blood storage on interferon-gamma production in cattle affected naturally with Mycobacterium bovis. Vet. Rec.160:660-662.","journal-title":"Vet. Rec."},{"key":"e_1_3_2_5_2","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1016\/j.rvsc.2005.11.005","volume":"81","year":"2006","unstructured":"de la Rua-Domenech, R., A. T. Goodchild, H. M. Vordermeier, R. G. Hewinson, K. H. Christiansen, and R. S. Clifton-Hadley. 2006. 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The effect of the tuberculin test and the consequences of a delay in blood culture on the sensitivity of a gamma-interferon assay for the detection of Mycobacterium bovis infection in cattle. Vet. Immunol. Immunopathol.102:413-420.","journal-title":"Vet. Immunol. Immunopathol."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1111\/j.1751-0813.1990.tb07730.x","volume":"67","year":"1990","unstructured":"Rothel, J. S., S. L. Jones, L. A. Corner, J. C. Cox, and P. R. Wood. 1990. A sandwich enzyme immunoassay for bovine interferon-gamma and its use for the detection of tuberculosis in cattle. Aust. Vet. J.67:134-137.","journal-title":"Aust. Vet. J."},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1053\/rvsc.2000.0386","volume":"69","year":"2000","unstructured":"Ryan, T. J., B. M. Buddle, and G. W. De Lisle. 2000. An evaluation of the gamma interferon test for detecting bovine tuberculosis in cattle 8 to 28 days after tuberculin skin testing. Res. Vet. Sci.69:57-61.","journal-title":"Res. Vet. Sci."},{"key":"e_1_3_2_11_2","unstructured":"Reference deleted."},{"key":"e_1_3_2_12_2","first-page":"72","volume":"16","year":"2006","unstructured":"Vordermeier, H. M., A. Whelan, K. Ewer, T. Goodchild, R. Clifton-Hadley, J. Williams, and R. G. Hewinson. 2006. The BOVIGAM\u00ae assay as ancillary test to the tuberculin skin test. Gov. Vet. J.16:72-80.","journal-title":"Gov. Vet. J."},{"key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1128\/CDLI.10.5.960-966.2003","volume":"10","year":"2003","unstructured":"Waters, W. R., M. V. Palmer, D. L. Whipple, M. P. Carlson, and B. J. Nonnecke. 2003. Diagnostic implications of antigen-induced gamma interferon, nitric oxide, and tumor necrosis factor alpha production by peripheral blood mononuclear cells from Mycobacterium bovis-infected cattle. Clin. Diagn. Lab. Immunol.10:960-966.","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1128\/CDLI.11.4.729-735.2004","volume":"11","year":"2004","unstructured":"Waters, W. R., B. J. Nonnecke, M. V. Palmer, S. Robbe-Austermann, J. P. Bannantine, J. R. Stabel, D. L. Whipple, J. B. Payeur, D. M. Estes, J. E. Pitzer, and F. C. Minion. 2004. Use of recombinant ESAT-6:CFP-10 fusion protein for differentiation of infections of cattle by Mycobacterium bovis and by M. avium subsp. avium and M. avium subsp. paratuberculosis. Clin. Diagn. Lab. Immunol.11:729-735.","journal-title":"Clin. Diagn. Lab. 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