{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T07:08:19Z","timestamp":1782457699322,"version":"3.54.5"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Research and Innovation Fund, National Science"},{"name":"Research and Innovation Grant, Ubon Ratchathani University"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p>Opisthorchiasis and cholangiocarcinoma (CCA) continue to be public health concerns in many Southeast Asian countries. Although the prevalence of opisthorchiasis is declining, reported cases tend to have a light-intensity infection. Therefore, early detection by using sensitive methods is necessary. Several sensitive methods have been developed to detect opisthorchiasis. The immunological detection of antigenic proteins has been proposed as a sensitive method for examining opisthorchiasis.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>The <jats:italic>Opisthorchis viverrini<\/jats:italic> antigenic proteins, including cathepsin B (OvCB), asparaginyl endopeptidase (OvAEP), and cathepsin F (OvCF), were used to construct multi-antigenic proteins. The protein sequences of OvCB, OvAEP, and OvCF, with a high probability of B cell epitopes, were selected using BepiPred 1.0 and the IEDB Analysis Resource. These protein fragments were combined to form OvCB_OvAEP_OvCF recombinant DNA, which was then used to produce a recombinant protein in <jats:italic>Escherichia coli<\/jats:italic> strain BL21(DE3). The potency of the recombinant protein as a diagnostic target for opisthorchiasis was assessed using immunoblotting and compared with that of the gold standard method, the modified formalin-ether concentration technique.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>The recombinant OvCB_OvAEP_OvCF protein showed strong reactivity with total immunoglobulin G (IgG) antibodies against light-intensity <jats:italic>O.\u00a0viverrini<\/jats:italic> infections in the endemic areas. Consequently, a high sensitivity (100%) for diagnosing opisthorchiasis was reported. However, cross-reactivity with sera from other helminth and protozoan infections (including taeniasis, strongyloidiasis, giardiasis, <jats:italic>E.\u00a0coli<\/jats:italic> infection, enterobiasis, and mixed infection of <jats:italic>Echinostome<\/jats:italic> spp. and <jats:italic>Taenia<\/jats:italic> spp.) and no reactivity with sera from patients with non-parasitic infections led to a reduced specificity of 78.4%. In addition, the false negative rate (FNR), false positive rate (FPR), positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy were 0%, 21.6%, 81.4%, 100%, and 88.9%, respectively.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>The high sensitivity of the recombinant OvCB_OvAEP_OvCF protein in detecting opisthorchiasis demonstrates its potential as an opisthorchiasis screening target. Nonetheless, research on reducing cross-reactivity should be undertaken by detecting other antibodies in other sample types, such as saliva, urine, and feces.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical Abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-024-06285-7","type":"journal-article","created":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T20:12:54Z","timestamp":1715112774000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Multi-epitope protein production and its application in the diagnosis of opisthorchiasis"],"prefix":"10.1186","volume":"17","author":[{"given":"Jittiyawadee","family":"Sripa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tarinee","family":"Chaiwong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,5,7]]},"reference":[{"key":"6285_CR1","doi-asserted-by":"crossref","unstructured":"Sithithaworn P, Andrews RH, Nguyen VD, Wongsaroj T, Sinuon M, Odermatt P, et\u00a0al. The current status of opisthorchiasis and clonorchiasis in the Mekong Basin. Parasitol Int. 2012;61:10\u20136.","DOI":"10.1016\/j.parint.2011.08.014"},{"key":"6285_CR2","doi-asserted-by":"crossref","unstructured":"Sayasone S, Utzinger J, Akkhavong K, Odermatt P. Multiparasitism and intensity of helminth infections in relation to symptoms and nutritional status among children: a cross-sectional study in southern Lao People\u2019s Democratic Republic. Acta Trop. 2015;141:322\u201331.","DOI":"10.1016\/j.actatropica.2014.09.015"},{"key":"6285_CR3","doi-asserted-by":"crossref","unstructured":"Miyamoto K, Kirinoki M, Matsuda H, Hayashi N, Chigusa Y, Sinuon M, et\u00a0al. Field survey focused on Opisthorchis viverrini infection in five provinces of Cambodia. Parasitol Int. 2014;63:366\u201373.","DOI":"10.1016\/j.parint.2013.12.003"},{"key":"6285_CR4","doi-asserted-by":"publisher","first-page":"838","DOI":"10.4269\/ajtmh.19-0864","volume":"102","author":"J Boondit","year":"2020","unstructured":"Boondit J, Suwannahitatorn P, Siripattanapipong S, Leelayoova S, Mungthin M, Tan-Ariya P, et\u00a0al. An epidemiological survey of Opisthorchis viverrini infection in a lightly infected community, Eastern Thailand. Am J Trop Med Hyg. 2020;102:838\u201343.","journal-title":"Am J Trop Med Hyg"},{"key":"6285_CR5","unstructured":"Jamornthanyawat N. The diagnosis of human opisthorchiasis. Southeast Asian J Trop Med Public Health. 2002;33:86\u201391."},{"key":"6285_CR6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0006030","volume":"11","author":"S Buathong","year":"2017","unstructured":"Buathong S, Leelayoova S, Mungthin M, Ruang-Areerate T, Naaglor T, Suwannahitatorn P, et\u00a0al. Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand. PLoS Negl Trop Dis. 2017;11:e0006030.","journal-title":"PLoS Negl Trop Dis"},{"key":"6285_CR7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007186","volume":"13","author":"C Worasith","year":"2019","unstructured":"Worasith C, Wangboon C, Duenngai K, Kiatsopit N, Kopolrat K, Techasen A, et\u00a0al. Comparing the performance of urine and copro-antigen detection in evaluating Opisthorchis viverrini infection in communities with different transmission levels in Northeast Thailand. PLoS Negl Trop Dis. 2019;13:e0007186.","journal-title":"PLoS Negl Trop Dis"},{"key":"6285_CR8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0004157","volume":"9","author":"C Worasith","year":"2015","unstructured":"Worasith C, Kamamia C, Yakovleva A, Duenngai K, Wangboon C, Sithithaworn J, et\u00a0al. Advances in the diagnosis of human Opisthorchiasis: development of Opisthorchis viverrini antigen detection in urine. PLoS Negl Trop Dis. 2015;9:e0004157.","journal-title":"PLoS Negl Trop Dis"},{"key":"6285_CR9","doi-asserted-by":"publisher","first-page":"196","DOI":"10.1016\/j.parint.2011.06.009","volume":"61","author":"P Sawangsoda","year":"2012","unstructured":"Sawangsoda P, Sithithaworn J, Tesana S, Pinlaor S, Boonmars T, Mairiang E, et\u00a0al. Diagnostic values of parasite-specific antibody detections in saliva and urine in comparison with serum in opisthorchiasis. Parasitol Int. 2012;61:196\u2013202.","journal-title":"Parasitol Int"},{"key":"6285_CR10","first-page":"593","volume":"28","author":"D Watthanakulpanich","year":"1997","unstructured":"Watthanakulpanich D, Waikagul J, Anantaphruti MT, Dekumyoy P. Evaluation of Bithynia funiculata snail antigens by ELISA-serodiagnosis of human opisthorchiasis. Southeast Asian J Trop Med Public Health. 1997;28:593\u20138.","journal-title":"Southeast Asian J Trop Med Public Health"},{"key":"6285_CR11","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1016\/S1383-5769(02)00013-2","volume":"51","author":"J Waikagul","year":"2002","unstructured":"Waikagul J, Dekumyoy P, Chaichana K, Thairungroje Anantapruti M, Komalamisra C, et\u00a0al. Serodiagnosis of human opisthorchiasis using cocktail and electroeluted Bithynia snail antigens. Parasitol Int. 2002;51:237\u201347.","journal-title":"Parasitol Int"},{"key":"6285_CR12","first-page":"813","volume":"41","author":"W Pakdee","year":"2010","unstructured":"Pakdee W, Waikagul J, Kalambaheti T, Ito A, Dekumyoy P. Iso-electricfocusing of Bithynia snail antigens for IgG- and IgG(1\u20134)-ELISA detection of human opisthorchiasis. Southeast Asian J Trop Med Public Health. 2010;41:813\u201320.","journal-title":"Southeast Asian J Trop Med Public Health"},{"key":"6285_CR13","first-page":"419","volume":"41","author":"A Chanawong","year":"1990","unstructured":"Chanawong A, Waikagul J, Thammapalerd N. Detection of shared antigens of human liver flukes Opisthorchis viverrini and its snail host. Bithynia spp. Trop Med Parasitol. 1990;41:419\u201321.","journal-title":"Bithynia spp Trop Med Parasitol"},{"key":"6285_CR14","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1016\/j.parint.2011.05.009","volume":"61","author":"J Sripa","year":"2012","unstructured":"Sripa J, Brindley PJ, Sripa B, Loukas A, Kaewkes S, Laha T. Evaluation of liver fluke recombinant cathepsin B-1 protease as a serodiagnostic antigen for human opisthorchiasis. Parasitol Int. 2012;61:191\u20135.","journal-title":"Parasitol Int"},{"key":"6285_CR15","doi-asserted-by":"publisher","first-page":"4571","DOI":"10.1007\/s00436-015-4703-9","volume":"114","author":"S Teimoori","year":"2015","unstructured":"Teimoori S, Arimatsu Y, Laha T, Kaewkes S, Sereerak P, Tangkawattana S, et\u00a0al. Immunodiagnosis of opisthorchiasis using parasite cathepsin F. Parasitol Res. 2015;114:4571\u20138.","journal-title":"Parasitol Res"},{"key":"6285_CR16","doi-asserted-by":"publisher","first-page":"e49","DOI":"10.1016\/j.ijid.2008.03.033","volume":"12","author":"T Laha","year":"2008","unstructured":"Laha T, Sripa J, Sripa B, Pearson M, Tribolet L, Kaewkes S, et\u00a0al. Asparaginyl endopeptidase from the carcinogenic liver fluke, Opisthorchis viverrini, and its potential for serodiagnosis. Int J Infect Dis. 2008;12:e49-59.","journal-title":"Int J Infect Dis"},{"key":"6285_CR17","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1186\/s13071-016-1418-4","volume":"9","author":"C Lv","year":"2016","unstructured":"Lv C, Hong Y, Fu Z, Lu K, Cao X, Wang T, et\u00a0al. Evaluation of recombinant multi-epitope proteins for diagnosis of goat schistosomiasis by enzyme-linked immunosorbent assay. Parasit Vectors. 2016;9:135.","journal-title":"Parasit Vectors"},{"key":"6285_CR18","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12879-016-1427-0","volume":"16","author":"M Almeida","year":"2016","unstructured":"Almeida M, Pizzini C, Damasceno L, Muniz M, Paes R, Peralta R, et\u00a0al. Validation of western blot for Histoplasma capsulatum antibody detection assay. BMC Infect Dis. 2016;16:1\u201318.","journal-title":"BMC Infect Dis"},{"key":"6285_CR19","doi-asserted-by":"publisher","first-page":"13370","DOI":"10.1038\/srep13370","volume":"5","author":"Y Guan","year":"2015","unstructured":"Guan Y, Zhu Q, Huang D, Zhao S, Jan Lo L, Peng J. An equation to estimate the difference between theoretically predicted and SDS PAGE-displayed molecular weights for an acidic peptide. Sci Rep. 2015;5:13370.","journal-title":"Sci Rep"},{"key":"6285_CR20","doi-asserted-by":"publisher","first-page":"356","DOI":"10.4269\/ajtmh.1988.38.356","volume":"38","author":"S Wongratanacheewin","year":"1988","unstructured":"Wongratanacheewin S, Bunnag D, Vaeusorn N, Sirisinha S. Characterization of humoral immune response in the serum and bile of patients with opisthorchiasis and its application in immunodiagnosis. Am J Trop Med Hyg. 1988;38:356\u201362.","journal-title":"Am J Trop Med Hyg"},{"key":"6285_CR21","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1111\/j.1365-2249.1991.tb08151.x","volume":"84","author":"MR Haswell-Elkins","year":"1991","unstructured":"Haswell-Elkins MR, Sithithaworn P, Mairiang E, Elkins DB, Wongratanacheewin S, Kaewkes S, et\u00a0al. Immune responsiveness and parasite-specific antibody levels in human hepatobiliary disease associated with Opisthorchis viverrini infection. Clin Exp Immunol. 1991;84:213\u20138.","journal-title":"Clin Exp Immunol"},{"key":"6285_CR22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000398","volume":"3","author":"P Pinlaor","year":"2009","unstructured":"Pinlaor P, Kaewpitoon N, Laha T, Sripa B, Kaewkes S, Morales ME, et\u00a0al. Cathepsin F cysteine protease of the human liver fluke. Opisthorchis viverrini.\u00a0PLoS Negl Trop Dis. 2009;3:e398.","journal-title":"Opisthorchis viverrini PLoS Negl Trop Dis"},{"key":"6285_CR23","doi-asserted-by":"publisher","first-page":"971","DOI":"10.1016\/0020-7519(94)00212-7","volume":"25","author":"PS Akai","year":"1995","unstructured":"Akai PS, Pungpak S, Chaicumpa W, Kitikoon V, Ruangkunaporn Y, Bunnag D, et\u00a0al. Serum antibody responses in opisthorchiasis. Int J Parasitol. 1995;25:971\u20133.","journal-title":"Int J Parasitol"},{"key":"6285_CR24","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1186\/s13071-022-05338-z","volume":"15","author":"KY Kopolrat","year":"2022","unstructured":"Kopolrat KY, Singthong S, Khuntikeo N, Loilome W, Worasith C, Homwong C, et\u00a0al. Performance of Mini Parasep\u00ae SF stool concentrator kit, Kato-Katz, and formalin-ethyl acetate concentration methods for diagnosis of opisthorchiasis in Northeast Thailand. Parasit Vectors. 2022;15:234.","journal-title":"Parasit Vectors"},{"key":"6285_CR25","doi-asserted-by":"publisher","first-page":"1403","DOI":"10.1093\/trstmh\/trab066","volume":"115","author":"W Phupiewkham","year":"2021","unstructured":"Phupiewkham W, Rodpai R, Inthavongsack S, Laymanivong S, Thanchomnang T, Sadaow L, et\u00a0al. High prevalence of opisthorchiasis in rural populations from Khammouane Province, central Lao PDR: serological screening using total IgG- and IgG4-based ELISA. Trans R Soc Trop Med Hyg. 2021;115:1403\u20139.","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"6285_CR26","doi-asserted-by":"publisher","first-page":"10879","DOI":"10.1021\/nn404325t","volume":"7","author":"JH Lee","year":"2013","unstructured":"Lee JH, Seo HS, Song JA, Kwon KC, Lee EJ, Kim HJ, et\u00a0al. Proteinticle engineering for accurate 3D diagnosis. ACS Nano. 2013;7:10879\u201386.","journal-title":"ACS Nano"},{"key":"6285_CR27","doi-asserted-by":"publisher","first-page":"416","DOI":"10.1016\/j.coche.2013.08.001","volume":"2","author":"D Blackstock","year":"2013","unstructured":"Blackstock D, Park M, Sun Q, Tsai S-L, Chen W. Engineering protein modules for diagnostic applications. Curr Opin Chem Eng. 2013;2:416\u201324.","journal-title":"Curr Opin Chem Eng"}],"container-title":["Parasites &amp; Vectors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-024-06285-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13071-024-06285-7\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-024-06285-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T20:13:08Z","timestamp":1715112788000},"score":1,"resource":{"primary":{"URL":"https:\/\/parasitesandvectors.biomedcentral.com\/articles\/10.1186\/s13071-024-06285-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,7]]},"references-count":27,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["6285"],"URL":"https:\/\/doi.org\/10.1186\/s13071-024-06285-7","relation":{},"ISSN":["1756-3305"],"issn-type":[{"value":"1756-3305","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,7]]},"assertion":[{"value":"17 January 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 April 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 May 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study adhered to the guidelines of the Declaration of Helsinki. All procedures involved in participants in this study were approved by the Human Ethics Committee of Ubon Ratchathani University (UBU-REC-46\u20132563).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent for participation"}},{"value":"All authors have read and agreed to the published version of the manuscript.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"206"}}