{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T22:06:20Z","timestamp":1783721180851,"version":"3.55.0"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,8,16]],"date-time":"2023-08-16T00:00:00Z","timestamp":1692144000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>Ovine babesiosis caused by <jats:italic>Babesia ovis<\/jats:italic> is an economically significant disease. Recently, a few <jats:italic>B. ovis<\/jats:italic>-specific proteins, including recombinant <jats:italic>B. ovis<\/jats:italic> secreted antigen-1 (rBoSA1), have been identified. Immunological analyses revealed that rBoSA1 resides within the cytoplasm of infected erythrocytes and exhibits robust antigenic properties for detecting anti-<jats:italic>B. ovis<\/jats:italic> antibodies. This protein is released into the bloodstream during the parasite\u2019s development. It would be possible to diagnose active infections by detecting this secretory protein. For this purpose, a rBoSA1-specific polyclonal antibody-based sandwich ELISA was optimized in this study. Blood samples taken from the naturally (n: 100) and experimentally (n: 15) infected sheep were analyzed for the presence of native BoSA1. The results showed that native BoSA1 was detectable in 98% of naturally infected animals. There was a positive correlation between parasitemia level in microscopy and protein density in sandwich ELISA. Experimentally infected animals showed positive reactions from the first or second day of inoculations. However, experimental infections carried out by <jats:italic>Rhipicephalus bursa<\/jats:italic> ticks revealed the native BoSA1 was detectable from the 7<jats:sup>th<\/jats:sup> day of tick attachment when the parasite began to be seen microscopically. Sandwich ELISA was sensitive enough to detect rBoSA1 protein at a 1.52 ng\/ml concentration. Additionally, no serological cross-reactivity was observed between animals infected with various piroplasm species, including <jats:italic>Babesia bovis<\/jats:italic>, <jats:italic>B. bigemina<\/jats:italic>, <jats:italic>B. caballi, B. canis, B. gibsoni, Theileria equi<\/jats:italic>, and <jats:italic>T. annulata.<\/jats:italic> Taken collectively, the findings show that the rBoSA1-specific polyclonal antibody-based sandwich ELISA can be successfully used to diagnose clinical <jats:italic>B. ovis<\/jats:italic> infections in sheep at the early stage.<\/jats:p>","DOI":"10.3389\/fcimb.2023.1238369","type":"journal-article","created":{"date-parts":[[2023,8,17]],"date-time":"2023-08-17T13:33:11Z","timestamp":1692279191000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Babesia ovis secreted antigen-1 is a diagnostic marker during the active Babesia ovis infections in sheep"],"prefix":"10.3389","volume":"13","author":[{"given":"Ferda","family":"Sevinc","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mo","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinuo","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Onur","family":"Ceylan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehmet Can","family":"Ulucesme","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sezayi","family":"Ozubek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Munir","family":"Aktas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuenan","family":"Xuan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,8,16]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1016\/j.vetpar.2005.05.057","article-title":"Development of a polymerase chain reaction method for diagnosis of Babesia ovis infection in sheep and goats","volume":"133","author":"Aktas","year":"2005","journal-title":"Vet. Parasitol."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.3791\/57994","article-title":"A versatile model of hard tick infestation on laboratory rabbits","volume":"140","author":"Almaz\u00e1n","year":"2018","journal-title":"J. Vis. Exp."},{"key":"B3","doi-asserted-by":"publisher","first-page":"S16","DOI":"10.1007\/s00436-001-0563-6","article-title":"Isolation and preliminary characterization of a large Babesia sp. from sheep and goats in the eastern part of Gansu Province, China","volume":"88","author":"Bai","year":"2002","journal-title":"Parasitol. Res."},{"key":"B4","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1017\/S0031182004005190","article-title":"Babesiosis of cattle","volume":"129","author":"Bock","year":"2004","journal-title":"Parasitology"},{"key":"B5","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1517\/14712598.4.4.551","article-title":"Development of secreted proteins as biotherapeutic agents","volume":"4","author":"Bonin-Debs","year":"2004","journal-title":"Expert Opin. Biol. Ther."},{"key":"B6","doi-asserted-by":"publisher","first-page":"3771","DOI":"10.1128\/JCM.40.10.3771-3775.2002","article-title":"Evaluation of an enzyme-linked immunosorbent assay with recombinant rhoptry-associated protein 1 antigen against Babesia bovis for the detection of specific antibodies in cattle","volume":"40","author":"Boonchit","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"648","DOI":"10.1007\/BF00931081","article-title":"An improved ELISA for the detection of antibodies against Babesia bovis using either a native or a recombinant B. bovis antigen","volume":"76","author":"Bose","year":"1990","journal-title":"Parasitol. Res."},{"key":"B8","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1016\/0304-4017(94)03111-9","article-title":"Current state and future trends in the diagnosis of babesiosis","volume":"57","author":"Bose","year":"1995","journal-title":"Vet. Parasitol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"162","DOI":"10.3390\/pathogens10020162","article-title":"Tick-borne hemoparasites of sheep: A molecular research in Turkey","volume":"10","author":"Ceylan","year":"2021","journal-title":"Pathogens"},{"key":"B10","first-page":"114","article-title":"Immunoreactive proteins: their production and importance in haemoprotozoan diseases","volume-title":"Animal Health Perspectives","author":"Ceylan","year":"2022"},{"key":"B11","doi-asserted-by":"publisher","first-page":"109034","DOI":"10.1016\/j.vetpar.2020.109034","article-title":"Endemic instability of ovine babesiosis in Turkey: A country-wide seroepidemiological study","volume":"278","author":"Ceylan","year":"2020","journal-title":"Vet. Parasitol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"231","DOI":"10.3390\/pathogens10020231","article-title":"Primary tick-borne protozoan and rickettsial infections of animals in Turkey","volume":"10","author":"Ceylan","year":"2021","journal-title":"Pathogens"},{"key":"B13","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1111\/j.1348-0421.2008.00020.x","article-title":"Evaluation of a rapid ELISA technique for detection of circulating antigens of Toxoplasma gondii","volume":"52","author":"Chen","year":"2008","journal-title":"Microbiol. Immunol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1186\/s13071-017-2016-9","article-title":"A novel modified-indirect ELISA based on spherical body protein 4 for detecting antibody during acute and long-term infections with diverse Babesia bovis strains","volume":"10","author":"Chung","year":"2017","journal-title":"Parasit. Vectors"},{"key":"B15","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1179\/000349804225003181","article-title":"Quantitative determination of sporozoites and circumsporozoite antigen in mosquitoes infected with Plasmodium falciparum or P. vivax","volume":"98","author":"De Arruda","year":"2004","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"B16","first-page":"412","article-title":"Bovine babesiosis","volume-title":"Manual of standards for diagnostic tests and vaccines","author":"De Vos","year":"2000"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.0020204","article-title":"Estimation of the total parasite biomass in acute falciparum malaria from plasma PfHRP2","volume":"2","author":"Dondorp","year":"2005","journal-title":"PloS Med."},{"key":"B18","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.ttbdis.2015.07.017","article-title":"Transmission of Babesia ovis by different Rhipicephalus bursa developmental stages and infected blood injection","volume":"7","author":"Erster","year":"2016","journal-title":"Ticks Tick Borne Dis."},{"key":"B19","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1007\/s11250-011-0023-y","article-title":"An epidemiological survey on bovine and ovine babesiosis in Kurdistan Province, western Iran","volume":"44","author":"Fakhar","year":"2012","journal-title":"Trop. Anim. Health Prod."},{"key":"B20","doi-asserted-by":"publisher","first-page":"2603","DOI":"10.1128\/JCM.39.7.2603-2609.2001","article-title":"Identification and expression of a 50-kilodalton surface antigen of Babesia gibsoni and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay","volume":"39","author":"Fukumoto","year":"2001","journal-title":"J. Clin. Microbiol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1016\/S0304-4017(01)00488-5","article-title":"Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily","volume":"99","author":"Georges","year":"2001","journal-title":"Vet. Parasitol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1316","DOI":"10.1128\/CVI.00150-08","article-title":"Validation of a competitive enzyme-linked immunosorbent assay for detection of Babesia bigemina antibodies in cattle","volume":"15","author":"Goff","year":"2008","journal-title":"Clin. Vaccine Immunol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1128\/CDLI.10.1.38-43.2003","article-title":"Competitive enzyme-linked immunosorbent assay based on a rhoptry-associated protein 1 epitope specifically identifies Babesia bovis-infected cattle","volume":"10","author":"Goff","year":"2003","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1212","DOI":"10.1128\/CVI.00196-06","article-title":"Validation of a competitive enzyme-linked immunosorbent assay for detection of antibodies against Babesia bovis","volume":"13","author":"Goff","year":"2006","journal-title":"Clin. Vaccine Immunol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1016\/j.exppara.2007.11.010","article-title":"Babesia gibsoni: serodiagnosis of infection in dogs by an enzyme-linked immunosorbent assay with recombinant BgTRAP","volume":"118","author":"Goo","year":"2008","journal-title":"Exp. Parasitol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1016\/j.exppara.2009.05.001","article-title":"A new ovine Babesia species transmitted by Hyalomma anatolicum","volume":"122","author":"Guan","year":"2009","journal-title":"Exp. Parasitol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1128\/CMR.13.3.451","article-title":"Babesiosis","volume":"13","author":"Homer","year":"2000","journal-title":"Clin. Microbiol. Rev."},{"key":"B28","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.vetpar.2006.03.013","article-title":"Evaluation of enzyme-linked immunosorbent assays with recombinant antigens for the serodiagnosis of equine Babesia infections","volume":"140","author":"Huang","year":"2006","journal-title":"Vet. Parasitol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"633","DOI":"10.1016\/S0020-7519(00)00008-4","article-title":"Detection of Babesia caballi infection by enzyme-linked immunosorbent assay using recombinant 48-kDa merozoite rhoptry protein","volume":"30","author":"Ikadai","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1051\/parasite\/2011183229","article-title":"A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan)","volume":"18","author":"Iqbal","year":"2011","journal-title":"Parasite"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1708","DOI":"10.3390\/foods10081708","article-title":"Non-specific binding and cross-reaction of ELISA: a case study of porcine hemoglobin detection","volume":"10","author":"Jiang","year":"2021","journal-title":"Foods"},{"key":"B32","first-page":"828","article-title":"Detection of P. vivax antigens in malaria endemic populations of Nepal by Elisa using monoclonal antibodies raised against Thai isolates","volume":"35","author":"Joshi","year":"2004","journal-title":"South Asian J. Trop. Med. Public Health"},{"key":"B33","doi-asserted-by":"publisher","first-page":"2285","DOI":"10.1128\/JCM.37.7.2285-2290.2999","article-title":"Detection of equine antibodies to Babesia caballi by recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay","volume":"37","author":"Kappmeyer","year":"1999","journal-title":"J. Clin. Microbiol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1016\/S0304-4017(02)00161-9","article-title":"Identification of immunoreactive polypeptides of Babesia equi parasite during immunization","volume":"107","author":"Kumar","year":"2002","journal-title":"Vet. Parasitol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1016\/j.exppara.2011.10.012","article-title":"A double antibody sandwich enzyme-linked immunosorbent assay for detection of secreted antigen 1 of Babesia microti using hamster model","volume":"130","author":"Luo","year":"2012","journal-title":"Exp. Parasitol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1648","DOI":"10.1128\/jcm.25.9.1648-1652.1987","article-title":"Detection of culture-derived Babesia bovis exoantigen using a two-site enzyme immunoassay","volume":"25","author":"Montealegre","year":"1987","journal-title":"J. Clin. Microbiol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"1990","DOI":"10.1017\/S0031182016001293","article-title":"Expression of sheep pathogen Babesia sp. Xinjiang rhoptry-associated protein 1 and evaluation of its diagnostic potential by enzyme-linked immunosorbent assay","volume":"143","author":"Niu","year":"2016","journal-title":"Parasitology"},{"key":"B38","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1016\/j.exppara.2010.06.018","article-title":"Babesia microti: molecular and antigenic characterizations of a novel 94-kDa protein (BmP94)","volume":"127","author":"Ooka","year":"2011","journal-title":"Exp. Parasitol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"13","DOI":"10.3390\/vetsci5010013","article-title":"Development of an indirect ELISA based on a recombinant chimeric protein for the detection of antibodies against bovine babesiosis","volume":"5","author":"Ortiz","year":"2018","journal-title":"Vet. Sci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"212","DOI":"10.1093\/jme\/tjw134","article-title":"Molecular and parasitological survey of ovine piroplasmosis, including the first report of Theileria annulata (Apicomplexa: Theileridae) in Sheep and Goats from Turkey","volume":"54","author":"Ozubek","year":"2017","journal-title":"J. Med. Entomol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"113","DOI":"10.3390\/pathogens12010113","article-title":"Discovery of a novel species infecting goats: morphological and molecular characterization of Babesia aktasi n. sp","volume":"12","author":"Ozubek","year":"2023","journal-title":"Pathogens"},{"key":"B42","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1590\/S0036-46651992000200010","article-title":"Evaluation of an ELISA technique for detection of antigens and circulating immune complexes of Trypanosoma cruzi by a field study in an endemic zone of Argentina","volume":"34","author":"Petray","year":"1992","journal-title":"Rev. Inst Med. Trop. Sao Paulo"},{"key":"B43","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1590\/S0074-02762009000700010","article-title":"Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis","volume":"104","author":"Ramos","year":"2009","journal-title":"Mem Inst Oswaldo Cruz"},{"key":"B44","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1007\/s00436-011-2658-z","article-title":"Babesia ovis as the main causative agent of sheep babesiosis in Iran","volume":"110","author":"Ranjbar-Bahadori","year":"2012","journal-title":"Parasitol. Res."},{"key":"B45","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1111\/j.1600-0854.2008.00723.x","article-title":"Secretion of proteins into host cells by apicomplexan parasites","volume":"9","author":"Ravindran","year":"2008","journal-title":"Traffic"},{"key":"B46","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/S0020-7519(99)00161-7","article-title":"A common high molecular weight antigen of Babesia bovis isolates from Mexico","volume":"30","author":"Sahagun Ruiz","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1128\/JCM.03219-14","article-title":"Identification and expression of Babesia ovis secreted antigen 1 and an evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay","volume":"53","author":"Sevinc","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"B48","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.vetpar.2012.07.025","article-title":"Babesia ovis infections: detailed clinical and laboratory observations in the pre- and post-treatment periods of 97 field cases","volume":"191","author":"Sevinc","year":"2013","journal-title":"Vet. Parasitol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.smallrumres.2013.10.010","article-title":"The effect of 12 successive blood passages on the virulence of Babesia ovis in splenectomized lambs: a preliminary study","volume":"116","author":"Sevinc","year":"2014","journal-title":"Small Rumin. Res."},{"key":"B50","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1016\/j.vetpar.2007.07.014","article-title":"Therapeutic and prophylactic efficacy of imidocarb dipropionate on experimental Babesia ovis infection of lambs","volume":"149","author":"Sevinc","year":"2007","journal-title":"Vet. Parasitol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/j.vetpar.2018.02.013","article-title":"Haemoparasitic agents associated with ovine babesiosis: a possible negative interaction between Babesia ovis and Theileria ovis","volume":"252","author":"Sevinc","year":"2018","journal-title":"Vet. Parasitol."},{"key":"B52","doi-asserted-by":"publisher","first-page":"101893","DOI":"10.1016\/j.cimid.2022.101893","article-title":"The presence of malignant ovine babesiosis in Bosnia and Herzegovina indicates a possible emerging risk for Balkan region","author":"Stevanovic","year":"2022","journal-title":"Comp. Immunol. Microbiol. Infect. Dis."},{"key":"B53","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1007\/s00436-007-0718-1","article-title":"A shared antigen among Babesia species: ribosomal phosphoprotein P0 as a universal babesial vaccine candidate","volume":"102","author":"Terkawi","year":"2007","journal-title":"Parasitol. Res."},{"key":"B54","doi-asserted-by":"publisher","first-page":"e00067","DOI":"10.1128\/JCM.00067-18","article-title":"BmGPAC, an antigen capture assay for detection of active Babesia microti infection","volume":"56","author":"Thekkiniath","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/j.vetpar.2006.01.035","article-title":"Babesia-a historical overview","volume":"138","author":"Uilenberg","year":"2006","journal-title":"Vet. Parasitol."},{"key":"B56","doi-asserted-by":"publisher","first-page":"514","DOI":"10.3390\/pathogens12040514","article-title":"Small ruminant piroplasmosis: High prevalence of Babesia aktasi n. sp. in goats in T\u00fcrkiye","volume":"12","author":"Ulucesme","year":"2023","journal-title":"Pathogens"},{"key":"B57","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1111\/j.1439-0450.1998.tb00807.x","article-title":"Clinical, clino-pathological and serological studies of Babesia ovis in experimentally infected sheep","volume":"45","author":"Yeruham","year":"1998","journal-title":"Zentralbl Veterinarmed B."},{"key":"B58","doi-asserted-by":"publisher","first-page":"11S","DOI":"10.1007\/BF02632908","article-title":"Babesiosis in China","volume":"29","author":"Yin","year":"1997","journal-title":"Trop. Anim. Health Prod."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1238369\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,17]],"date-time":"2023-08-17T13:33:14Z","timestamp":1692279194000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1238369\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,16]]},"references-count":58,"alternative-id":["10.3389\/fcimb.2023.1238369"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1238369","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8,16]]},"article-number":"1238369"}}