{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T22:34:09Z","timestamp":1776897249739,"version":"3.51.2"},"reference-count":46,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2009,10,1]],"date-time":"2009-10-01T00:00:00Z","timestamp":1254355200000},"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":["J Clin Microbiol"],"published-print":{"date-parts":[[2009,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Brucellosis is a globally significant zoonosis, the control of which is difficult and resource intensive. Serological tests form a vital part of a multifactorial approach to control and are often performed in large numbers. The aim of the present study was to develop a new assay to improve the efficiency, ease, and effectiveness of serological testing. An existing competitive enzyme-linked immunosorbent assay (cELISA) was adapted to a completely homogeneous time-resolved fluorescent resonance energy transfer (TR-FRET) assay. This was achieved by labeling an anti-<jats:italic>Brucella<\/jats:italic>monoclonal antibody with a long-lifetime donor fluorophore and<jats:italic>Brucella<\/jats:italic>smooth lipopolysaccharide with a compatible acceptor and optimizing the reading conditions. The assay was performed in a 96-well plate with a single 30-min incubation period and no separation (wash) steps and was concluded by a single plate-reading step. The performance of the assay was evaluated with a panel of serum samples from infected (<jats:italic>n<\/jats:italic>= 73) and uninfected (<jats:italic>n<\/jats:italic>= 480) sources and compared to the performance of the parent cELISA, an indirect ELISA (iELISA), and fluorescence polarization assay (FPA). The performance of the TR-FRET assay matched the performance of the iELISA, which had 100% diagnostic sensitivity and specificity, and surpassed the performance of the cELISA and the FPA. The results also demonstrated that the TR-FRET technique is effective with poor-quality serum samples from the field. To the knowledge of the authors, this is the first homogeneous TR-FRET assay to detect antibodies raised against an infectious disease. The technique appears to be sufficiently adaptable to meet the needs of many other similar testing requirements to identify infectious diseases.<\/jats:p>","DOI":"10.1128\/jcm.00919-09","type":"journal-article","created":{"date-parts":[[2009,8,6]],"date-time":"2009-08-06T01:10:45Z","timestamp":1249521045000},"page":"3098-3107","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":16,"title":["Time-Resolved Fluorescent Resonance Energy Transfer Assay for Simple and Rapid Detection of Anti-<i>Brucella<\/i>Antibodies in Ruminant Serum Samples"],"prefix":"10.1128","volume":"47","author":[{"given":"John A.","family":"McGiven","sequence":"first","affiliation":[{"name":"Brucella Research Group, Statutory and Exotic Bacteria, Veterinary Laboratories Agency, Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iain J.","family":"Thompson","sequence":"additional","affiliation":[{"name":"Brucella Research Group, Statutory and Exotic Bacteria, Veterinary Laboratories Agency, Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicola J.","family":"Commander","sequence":"additional","affiliation":[{"name":"Brucella Research Group, Statutory and Exotic Bacteria, Veterinary Laboratories Agency, Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judy A.","family":"Stack","sequence":"additional","affiliation":[{"name":"Brucella Research Group, Statutory and Exotic Bacteria, Veterinary Laboratories Agency, Weybridge, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1177\/1087057107303973"},{"key":"e_1_3_2_3_2","first-page":"207","volume":"12","year":"1999","unstructured":"Araj, G. F. 1999. Human brucellosis: a classical infectious disease with persistent diagnostic challenges. Clin. Lab. Sci.12:207-212.","journal-title":"Clin. Lab. Sci."},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/45.6.855"},{"key":"e_1_3_2_5_2","first-page":"148","year":"1965","unstructured":"Brownlee, K. A. 1965. Statistical theory and methodology in science and engineering, 2nd ed., p. 148-149. John Wiley & Sons, Inc., New York, NY.","journal-title":"Statistical theory and methodology in science and engineering, 2nd ed."},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.12.8353-8361.2005"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.2174\/138620703771826892"},{"key":"e_1_3_2_8_2","first-page":"927","volume":"55","year":"1985","unstructured":"Corbel, M. J. 1985. Recent advances in the study of Brucella antigens and their serological cross-reactions. Vet. Bull.55:927-942.","journal-title":"Vet. Bull."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1177\/108705710000500409","volume":"5","year":"2000","unstructured":"Enomoto, K., Y. Aono, T. Mitsugi, K. Takahashi, R. Suzuki, M. Pr\u00e9audat, G. Mathis, G. Kominami, and H. Takemoto. 2000. High throughput screening for human interferon-gamma production inhibitor using homogenous time-resolved fluorescence. J. Biomol. Screen.5:263-268.","journal-title":"J. Biomol. Screen."},{"key":"e_1_3_2_10_2","first-page":"55","volume":"2","year":"1948","unstructured":"F\u00f6rster, T. 1948. Zwischenmolekulare Energiewanderung und Fluoreszenz. Ann. Physik2:55-75.","journal-title":"Ann. Physik"},{"key":"e_1_3_2_11_2","first-page":"974","year":"2008","unstructured":"Garin-Bastuji, B., and J. M. Blasco. 2008. Caprine and ovine brucellosis (excluding Brucella ovis), p. 974-983. InManual of standards for diagnostic tests and vaccines. Office International des Epizooties, Paris, France.","journal-title":"Manual of standards for diagnostic tests and vaccines."},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(02)00217-1"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(95)00121-P"},{"key":"e_1_3_2_14_2","first-page":"616","volume":"32","year":"1985","unstructured":"Greiser-Wilke, I., V. Moennig, D. Thon, and K. Rauter. 1985. Characterization of monoclonal antibodies against Brucella melitensis. Zentralbl. Veterinarmed. B32:616-627.","journal-title":"Zentralbl. Veterinarmed. B"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2006.09.025"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja0452368"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(97)00202-2"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2004.12.003"},{"key":"e_1_3_2_19_2","first-page":"2821","volume":"31","year":"1997","unstructured":"Lin, M., and K. Nielsen. 1997. Binding of the Brucella abortus lipopolysaccharide O-chain fragment to a monoclonal antibody. Quantitative analysis by fluorescence quenching and polarization. J. Biol. Chem.31:2821-2827.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/39.9.1953"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jala.2006.09.003"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2007.03.003"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2008.05.004"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(03)00201-1"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2006.12.008"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1586\/14737159.5.4.561"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(88)90524-6"},{"key":"e_1_3_2_28_2","first-page":"177","volume":"2","year":"1999","unstructured":"Nasir, M. S., and M. E. Jolley. 1999. Fluorescence polarization: an analytical tool for immunoassay and drug discovery. Comb. Chem. High Throughput Screen.2:177-190.","journal-title":"Comb. Chem. High Throughput Screen."},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(96)00116-0"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(00)00386-2"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1081\/IAS-100104706"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1081\/IAS-100104705"},{"key":"e_1_3_2_33_2","first-page":"624","year":"2008","unstructured":"Nielsen, K., and D. R. Ewalt. 2008. Bovine brucellosis, p. 624-659. In Manual of standards for diagnostic tests and vaccines. Office International des Epizooties, Paris, France.","journal-title":"Office International des Epizooties, Paris, France"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(06)70382-6"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(02)00030-3"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.71.3.1481-1490.2003"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1373\/49.7.1105"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.2000.0338"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.21.10024"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1136\/vr.145.25.735"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.12.5426"},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1177\/1087057104268805"},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2005.11.052"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1515\/znb-1952-0303"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/cdli.3.3.309-314.1996"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.65.5.1939-1943.1997"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1177\/1087057105275628"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.00919-09","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.00919-09","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,26]],"date-time":"2023-05-26T07:45:43Z","timestamp":1685087143000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.00919-09"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,10]]},"references-count":46,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2009,10]]}},"alternative-id":["10.1128\/JCM.00919-09"],"URL":"https:\/\/doi.org\/10.1128\/jcm.00919-09","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,10]]},"assertion":[{"value":"2009-05-09","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2009-07-28","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2009-10-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}