{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:42:57Z","timestamp":1785415377537,"version":"3.56.0"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"European Union\u2019s Horizon 2020 research and innovation program","award":["101000467"],"award-info":[{"award-number":["101000467"]}]},{"name":"UGhent BOF funding","award":["bof.baf.2024.0712.01"],"award-info":[{"award-number":["bof.baf.2024.0712.01"]}]},{"name":"Strategic Research Program","award":["SRP47"],"award-info":[{"award-number":["SRP47"]}]},{"name":"Strategic Research Program","award":["SRP83"],"award-info":[{"award-number":["SRP83"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,12,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei<\/jats:italic>\n                      infections cause African trypanosomiasis, also known as sleeping sickness in humans and nagana in animals, presenting a significant global health and economic burden, especially in sub-Saharan Africa. Animal trypanosomiasis also affects the economic development in Asia and South America. Accurate diagnosis of diseases caused by infection with the parasite of the\n                      <jats:italic toggle=\"yes\">T. brucei<\/jats:italic>\n                      group remains a major challenge due to the persistence of infection-induced antibodies long after parasite clearance, complicating serological discrimination between active and past infections. To address this limitation, we developed a sensitive and specific antigen detection assay targeting\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei<\/jats:italic>\n                      enolase (\n                      <jats:italic toggle=\"yes\">Tbr<\/jats:italic>\n                      ENO) using a panel of camelid single-domain antibodies (sdAbs also known as nanobodies). Among the candidates, the sdAbsR3-77\/sdAbR2-103 sandwich enzyme-linked immunosorbent assay (ELISA) exhibited robust performance in detecting circulating\n                      <jats:italic toggle=\"yes\">Tbr<\/jats:italic>\n                      ENO in plasma from experimentally infected mice. Additionally, this assay showed strong potential as a \u201ctest-of-cure\u201d tool by monitoring real-time antigenemia throughout a chronic\n                      <jats:italic toggle=\"yes\">T. brucei<\/jats:italic>\n                      infection course. We further validated the assay\u2019s diagnostic utility in human clinical samples, detecting\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei rhodesiense<\/jats:italic>\n                      infections at both early and advanced stages and\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei gambiense<\/jats:italic>\n                      infections in advanced stage. The sdAbsR3-77H\/sdAbR2-103HA ELISA targeting\n                      <jats:italic toggle=\"yes\">Tbr<\/jats:italic>\n                      ENO shows potential for point-of-care pan-diagnosis of active\n                      <jats:italic toggle=\"yes\">T. brucei<\/jats:italic>\n                      infections (including\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei brucei<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">T. b. gambiense<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">T. b. rhodesiense<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei evansi<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">Trypanosoma brucei equiperdum<\/jats:italic>\n                      ) in both animals and humans. Therefore, this assay addresses gaps in current diagnostic capabilities by overcoming the key limitations of antibody-based tests, offering a promising tool for improved disease control.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        African trypanosomiasis, commonly known as sleeping sickness in humans and nagana in animals, is a life-threatening disease that remains a major health and economic concern in many parts of the world. One of the key difficulties in managing this disease is detecting ongoing infections, as existing antibody-based tests cannot reliably distinguish between current and past infections. In this study, we developed a novel laboratory test that detects a specific protein released by the parasite during infection. This test uses special antibodies derived from camels, known for their exceptional stability and precision, to accurately identify infections caused by multiple\n                        <jats:italic toggle=\"yes\">Trypanosoma brucei<\/jats:italic>\n                        subspecies. Our approach not only enables accurate diagnosis but also offers a way to monitor treatment success. This work provides a valuable tool for disease control efforts and could help improve the health of both humans and animals in regions where trypanosomiasis is endemic.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.00561-25","type":"journal-article","created":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T13:29:06Z","timestamp":1761830946000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Development of a camelid single-domain antibody-based antigen detection assay for the pan-specific diagnosis of active human and animal\n                    <i>Trypanosoma brucei<\/i>\n                    infections"],"prefix":"10.1128","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9368-5827","authenticated-orcid":true,"given":"Zeng","family":"Li","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo-kyung","family":"Jin","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily Estefania Timaury","family":"Moreno","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9s","family":"\u00c1lvarez-Rodr\u00edguez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]},{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biochemistry and Microbiology, Ghent University","place":["Ghent, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo A. Van","family":"Ginderachter","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]},{"id":[{"id":"https:\/\/ror.org\/04q4ydz28","id-type":"ROR","asserted-by":"publisher"}],"name":"Myeloid Cell Immunology Laboratory, VIB Center for Inflammation Research","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magdalena","family":"Radwanska","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory for Biomedical Research, Department of Environmental Technology, Food Technology and Molecular Biotechnology, Ghent University Global Campus","place":["Incheon, South Korea"]},{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomedical Molecular Biology, Ghent University","place":["Ghent, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7420-0983","authenticated-orcid":true,"given":"Yann G. -J.","family":"Sterckx","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/008x57b05","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Medical Biochemistry (LMB) and the Infla-Med Centre of Excellence, Department of Pharmaceutical Sciences, University of Antwerp","place":["Antwerp, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benoit","family":"Stijlemans","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]},{"id":[{"id":"https:\/\/ror.org\/04q4ydz28","id-type":"ROR","asserted-by":"publisher"}],"name":"Myeloid Cell Immunology Laboratory, VIB Center for Inflammation Research","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3760-7968","authenticated-orcid":false,"given":"Stefan","family":"Magez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006e5kg04","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratory of Cellular and Molecular Immunology, Brussels Center for Immunology (BCIM), Department of Bioengineering Sciences (DBIT), Vrije Universiteit Brussel (VUB)","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000961"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.2147\/CLEP.S39728"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1017\/s0031182000077805"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001857"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.2005.73.491"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa063265"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001454"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2017.02.008"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3156.2005.01470.x"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2014.02.043"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0304-4017(02)00270-4"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.12.5517-5524.2003"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008044"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms12010044"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(92)90174-b"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0304-4017(94)00721-n"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0020-7519(96)00160-9"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(92)90435-f"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004360050594"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0020-7519(98)00123-4"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12967-014-0352-5"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/363446a0"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0161-5890(03)00084-1"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/7.9.1129"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-26732-7"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines8030415"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4894(70)90120-7"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2022.09.005"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0004420"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.11.051"},{"key":"e_1_3_4_32_2","first-page":"792","article-title":"Duration of symptoms and case fatality of sleeping sickness caused by Trypanosoma brucei rhodesiense in Tororo, Uganda","volume":"74","author":"Odiit M","year":"1997","unstructured":"Odiit M, Kansiime F, Enyaru JC. 1997. Duration of symptoms and case fatality of sleeping sickness caused by Trypanosoma brucei rhodesiense in Tororo, Uganda. East Afr Med J 74:792\u2013795.","journal-title":"East Afr Med J"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000303"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2334-8-16"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.00561-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.00561-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T14:01:40Z","timestamp":1765980100000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jcm.00561-25"}},"subtitle":[],"editor":[{"given":"Bobbi S.","family":"Pritt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,12,17]]},"references-count":33,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2025,12,17]]}},"alternative-id":["10.1128\/jcm.00561-25"],"URL":"https:\/\/doi.org\/10.1128\/jcm.00561-25","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,17]]},"assertion":[{"value":"2025-04-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00561-25"}}