{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T07:25:59Z","timestamp":1790925959116,"version":"4.1.0"},"reference-count":23,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T00:00:00Z","timestamp":1772236800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T00:00:00Z","timestamp":1772236800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,3]]},"abstract":"Abstract<\/jats:title>\n \n Schistosomes are parasitic trematode worms of the genus\n Schistosoma<\/jats:italic>\n , responsible for causing urogenital and intestinal schistosomiasis. Six primary species infect humans:\n S. mansoni<\/jats:italic>\n ,\n S. japonicum<\/jats:italic>\n ,\n S. mekongi<\/jats:italic>\n ,\n S. intercalatum<\/jats:italic>\n ,\n S. guineensis<\/jats:italic>\n , and\n S. haematobium<\/jats:italic>\n . In addition, several species including\n S. bovis<\/jats:italic>\n ,\n S. curassoni<\/jats:italic>\n , and\n S. mattheei<\/jats:italic>\n primarily infect animals, particularly cattle. Hybridization events have been documented both between human\u2010infecting species and between human\u2010 and animal\u2010infecting species of\n Schistosoma<\/jats:italic>\n . Current methods for detecting hybrids rely on genotyping mitochondrial cytochrome oxidase I (cox1) and the ribosomal internal transcribed spacer (ITS), or whole\u2010genome sequencing. This protocol describes a multi\u2010locus, multiplex amplicon sequencing approach using Nanopore technology, referred to as NMAS\u2010Seq, for species\u2010level genotyping and hybrid detection. It includes comprehensive steps for multiplex nested PCR, amplicon quantification and pooling, library preparation, and sequencing. The workflow is adaptable for molecular typing of other parasitic species. \u00a9 2026 The Author(s).\n Current Protocols<\/jats:italic>\n published by Wiley Periodicals LLC.\n <\/jats:p>\n \n Basic Protocol 1<\/jats:bold>\n : DNA extraction and multiplex nested PCR\n <\/jats:p>\n \n Basic Protocol 2<\/jats:bold>\n : Amplicon quantification and pooling\n <\/jats:p>\n \n Basic Protocol 3<\/jats:bold>\n : Amplicon library preparation and Nanopore sequencing\n <\/jats:p>","DOI":"10.1002\/cpz1.70333","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T21:27:45Z","timestamp":1773005265000},"source":"Crossref","is-referenced-by-count":3,"title":["Schistosoma<\/i>\n Species and Hybrid Genotyping With a Field Deployable Multi\u2010Marker Amplicon Panel"],"prefix":"10.1002","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6576-0241","authenticated-orcid":false,"given":"Oluwaremilekun Grace","family":"Ajakaye","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Parasitology and Genomics of Neglected Tropical Diseases, Department of Animal and Environment Biology Adekunle Ajasin University Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael E.","family":"Grigg","sequence":"additional","affiliation":[{"name":"Molecular Parasitology Section, Laboratory of Parasitic Diseases, NIAID National Institutes of Health Bethesda Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,2,28]]},"reference":[{"key":"e_1_2_7_2_1_1","doi-asserted-by":"publisher","DOI":"10.3390\/tropicalmed6030109"},{"key":"e_1_2_7_2_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpara.2016.08.001"},{"key":"e_1_2_7_2_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140\u20106736(13)61949\u20102"},{"key":"e_1_2_7_2_4_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13071\u2010020\u201004476\u20108"},{"key":"e_1_2_7_2_5_1","first-page":"49","volume-title":"Amebiasis: Biology and pathogenesis of Entamoeba","author":"Gilchrist C. 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Encyclopedia.pub.https:\/\/encyclopedia.pub\/entry\/12259"},{"key":"e_1_2_7_2_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2021.104727"},{"key":"e_1_2_7_2_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2003.08.006"},{"key":"e_1_2_7_2_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00951\u201012"},{"key":"e_1_2_7_2_18_1","unstructured":"World Health Organization. (2020).Ending the neglect to attain the Sustainable Development Goals: A road map for neglected tropical diseases 2021\u20132030.https:\/\/www.who.int\/publications\/i\/item\/9789240010352"},{"key":"e_1_2_7_2_19_1","unstructured":"World Health Organization. (2025a). Schistosomiasis.https:\/\/www.who.int\/health\u2010topics\/schistosomiasis(Accessed May 24 2025)"},{"key":"e_1_2_7_2_20_1","unstructured":"World Health Organization. (2025b). Schistosomiasis.https:\/\/www.who.int\/news\u2010room\/fact\u2010sheets\/detail\/schistosomiasis(Accessed May 24 2025)"},{"key":"e_1_2_7_2_21_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001049"},{"key":"e_1_2_7_2_22_1","doi-asserted-by":"publisher","DOI":"10.5772\/intechopen.103680"},{"key":"e_1_2_7_3_2_1","unstructured":"https:\/\/www.promega.com\/resources\/tools\/biomath\/"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70333","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70333","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70333","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T02:15:40Z","timestamp":1789438540000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70333"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,28]]},"references-count":23,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,3]]}},"alternative-id":["10.1002\/cpz1.70333"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70333","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,28]]},"article-number":"e70333"}}