{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T17:48:15Z","timestamp":1783619295394,"version":"3.55.0"},"reference-count":29,"publisher":"SAGE Publications","issue":"3","license":[{"start":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T00:00:00Z","timestamp":1770422400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T00:00:00Z","timestamp":1770422400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"Equine Research Centre"},{"DOI":"10.13039\/501100001343","name":"university of pretoria","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001343","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J VET Diagn Invest"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:p>\n                    African horse sickness (AHS) is the only equine disease for which the World Organisation for Animal Health (WOAH) gives official disease-free status, given that it poses a major threat to the equine industry. The disease is caused by AHS virus (AHSV; family\n                    <jats:italic toggle=\"yes\">Sedoreoviridae<\/jats:italic>\n                    , taxon species\n                    <jats:italic toggle=\"yes\">Orbivirus alphaequi<\/jats:italic>\n                    ), which is endemic in sub-Saharan Africa. Reverse-transcription quantitative real-time PCR (RT-qPCR) is a rapid, sensitive detection method used in the diagnosis of AHS and the certification of animals as negative for AHSV for the purpose of movement. Genetic variability of AHSV may influence the accuracy of RT-qPCR detection methods because of possible mispriming and\/or probe binding failures. We evaluated the diagnostic accuracy of the current WOAH-recommended RT-qPCR assays for the detection of AHSV, namely the Ag\u00fcero et al. and Guthrie et al. methods. Utilizing 150 AHSV-positive diagnostic samples, we performed in vitro analysis using both assays. The Ag\u00fcero assay failed to detect AHSV in 13 samples (8.7% false-negative rate). The AHSV VP7 genes of the 13 negative samples, and publicly archived sequences were used to perform in silico analysis, and we incorporated minor changes into the primers and probes of modified Guthrie and modified Ag\u00fcero assays. A second in vitro analysis yielded 100% sensitivity for both assays. Differences in both the in silico and in vitro analyses highlight the need for continuous monitoring of the efficacy of molecular protocols used for the detection of AHSV.\n                  <\/jats:p>","DOI":"10.1177\/10406387261417355","type":"journal-article","created":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T10:36:25Z","timestamp":1770460585000},"page":"437-445","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":1,"title":["Enhancing African horse sickness virus detection: comparing and adapting PCR assays"],"prefix":"10.1177","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9176-0977","authenticated-orcid":false,"given":"Lisa","family":"Penzhorn","sequence":"first","affiliation":[{"name":"Equine Research Centre, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan E.","family":"Crafford","sequence":"additional","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan J.","family":"Guthrie","sequence":"additional","affiliation":[{"name":"Equine Research Centre, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2026,2,7]]},"reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063870802000310"},{"key":"e_1_3_2_3_2","first-page":"77","article-title":"The 1944 epizootic of horse-sickness in the Middle East","volume":"23","author":"Alexander RA","year":"1948","unstructured":"Alexander RA. The 1944 epizootic of horse-sickness in the Middle East. Onderstepoort J Vet Sci Anim Ind 1948;23:77\u201392.","journal-title":"Onderstepoort J Vet Sci Anim Ind"},{"key":"e_1_3_2_4_2","unstructured":"Arter-Hazzard M et al Bluetongue virus in Europe. Preliminary outbreak assessments qualitative risk assessments and updated situation assessments for bluetongue virus in Europe. Department for Environment Food and Rural Affairs 2025. https:\/\/www.gov.uk\/government\/publications\/bluetongue-virus-in-europe"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tree.2007.11.010"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2708.210004"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-ento-031616-035010"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/evj.13353"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/148.4.1667"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.12968"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jala.2007.05.001"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.05.044"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2012.12.014"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063879000200108"},{"key":"e_1_3_2_15_2","first-page":"329","article-title":"The 1960 epizootic of African horsesickness in the Middle East and SW Asia","volume":"31","author":"Howell PG","year":"1960","unstructured":"Howell PG. The 1960 epizootic of African horsesickness in the Middle East and SW Asia. J S Afr Vet Assoc 1960;31:329\u2013334.","journal-title":"J S Afr Vet Assoc"},{"key":"e_1_3_2_16_2","first-page":"139","article-title":"The isolation and identification of further antigenic types of African horsesickness virus","volume":"29","author":"Howell PG.","year":"1962","unstructured":"Howell PG. The isolation and identification of further antigenic types of African horsesickness virus. Onderstepoort J Vet Res 1962;29:139\u2013149.","journal-title":"Onderstepoort J Vet Res"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkf436"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/mst010"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bts199"},{"key":"e_1_3_2_20_2","first-page":"1764","article-title":"Outbreak of African horse sickness in Thailand, 2020","volume":"67","author":"King S","year":"2020","unstructured":"King S, et al Outbreak of African horse sickness in Thailand, 2020. Transbound Emerg Dis 2020;67:1764\u20131767.","journal-title":"Transbound Emerg Dis"},{"key":"e_1_3_2_21_2","first-page":"1751","article-title":"African horse sickness: its emergence in Thailand and potential threat to other Asian countries","volume":"67","author":"Lu G","year":"2020","unstructured":"Lu G, et al African horse sickness: its emergence in Thailand and potential threat to other Asian countries. Transbound Emerg Dis 2020;67:1751\u20131753.","journal-title":"Transbound Emerg Dis"},{"key":"e_1_3_2_22_2","first-page":"465","article-title":"Immunological types of horse sickness virus and their significance in immunization","volume":"27","author":"McIntosh BM.","year":"1958","unstructured":"McIntosh BM. Immunological types of horse sickness virus and their significance in immunization. Onderstepoort J Vet Res 1958;27:465\u2013532.","journal-title":"Onderstepoort J Vet Res"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1366"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2010.03.009"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1135(92)90041-Q"},{"key":"e_1_3_2_26_2","first-page":"311","article-title":"Validation of ELISA for the detection of African horse sickness virus antigens and antibodies","volume":"14","author":"Rubio C","year":"1998","unstructured":"Rubio C, et al Validation of ELISA for the detection of African horse sickness virus antigens and antibodies. Arch Virol Suppl 1998;14:311\u2013315.","journal-title":"Arch Virol Suppl"},{"key":"e_1_3_2_27_2","unstructured":"ThermoFisher. Tm Calculator for Primers. 2025. https:\/\/www.thermofisher.com\/ca\/en\/home\/brands\/thermo-scientific\/molecular-biology\/molecular-biology-learning-center\/molecular-biology-resource-library\/thermo-scientific-web-tools\/tm-calculator.html"},{"key":"e_1_3_2_28_2","unstructured":"World Organisation for Animal Health (WOAH). African horse sickness virus. The State of the World\u2019s Animal Health. WOAH 2023. https:\/\/www.woah.org\/en\/the-state-of-the-worlds-animal-health\/"},{"key":"e_1_3_2_29_2","unstructured":"World Organisation for Animal Health (WOAH). African horse sickness (infection with African horse sickness virus). In: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals [Internet]. WOAH 2025. https:\/\/www.woah.org\/fileadmin\/Home\/eng\/Health_standards\/tahm\/A_summry.htm"},{"key":"e_1_3_2_30_2","first-page":"385","article-title":"Diagnosis and molecular epidemiology of the African horsesickness virus by the polymerase chain reaction and restriction patterns","volume":"24","author":"Zientara S","year":"1993","unstructured":"Zientara S, et al Diagnosis and molecular epidemiology of the African horsesickness virus by the polymerase chain reaction and restriction patterns. Vet Res 1993;24:385\u2013395.","journal-title":"Vet Res"}],"container-title":["Journal of Veterinary Diagnostic Investigation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/10406387261417355","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1177\/10406387261417355","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/10406387261417355","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T00:29:49Z","timestamp":1777681789000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/10.1177\/10406387261417355"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,7]]},"references-count":29,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,5]]}},"alternative-id":["10.1177\/10406387261417355"],"URL":"https:\/\/doi.org\/10.1177\/10406387261417355","relation":{},"ISSN":["1040-6387","1943-4936"],"issn-type":[{"value":"1040-6387","type":"print"},{"value":"1943-4936","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,7]]}}}