{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T22:50:26Z","timestamp":1780527026388,"version":"3.54.1"},"reference-count":21,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2026,2,23]],"date-time":"2026-02-23T00:00:00Z","timestamp":1771804800000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T00:00:00Z","timestamp":1769904000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000061","name":"Fogarty International Center","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000061","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    African trypanosomes exhibit significant genetic diversity but share similar morphological characteristics, making effective surveillance and diagnosis challenging with conventional microscopy and other detection methods. While polymerase chain reaction (PCR) offers enhanced specificity and sensitivity, it requires DNA extraction, making it time\u2010consuming and costly, especially for resource\u2010limited regions such as ours (Africa). Additionally, amplification using DNA from the host's whole blood often fails due to the presence of polymerase inhibitors or low parasite DNA, particularly during chronic infections. This study introduces a novel PCR method that overcomes these challenges by using purified trypanosome cells as templates, bypassing the need for DNA extraction.\n                    <jats:italic>Trypanosoma brucei brucei<\/jats:italic>\n                    cells were isolated from mice blood using DEAE\u2013cellulose anion\u2010exchange chromatography and directly used in PCR amplification of the ITS\u20101 ribosomal RNA gene. Successful amplification was achieved with as few as 1.25\u00a0\u00d7\u00a010\u2075 isolated trypanosome cells, while gDNA extracted from parasitized blood failed to amplify. This cost\u2010effective and highly sensitive method improves species differentiation and enhances diagnostic capabilities, especially in cases of low parasitemia. The approach represents a significant advancement in trypanosome diagnosis, offering a faster, affordable, and reliable alternative to traditional methods. These findings support improved epidemiological surveillance and may contribute to the World Health Organization's goal of eliminating African trypanosomiasis, a disease that continues to present major economic and public health challenges in Africa. A graphical abstract of this study is presented in Figure 1. \u00a9 2026 Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Isolation of\n                    <jats:italic>Trypanosoma brucei brucei<\/jats:italic>\n                    from parasitized blood\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : PCR characterization of isolated\n                    <jats:italic>T. brucei brucei<\/jats:italic>\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70327","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T23:44:55Z","timestamp":1773013495000},"source":"Crossref","is-referenced-by-count":1,"title":["A Novel Cell\u2010Based PCR Method for Accurate Characterization of African Trypanosomes"],"prefix":"10.1002","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2092-0045","authenticated-orcid":false,"given":"Emmanuel Oluwadare","family":"Balogun","sequence":"first","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Kaduna Nigeria"},{"name":"Africa Center of Excellence for Neglected Tropical Diseases and Forensic Biotechnology (ACENTDFB) Ahmadu Bello University  Zaria Kaduna Nigeria"},{"name":"Department of Biomedical Chemistry, Graduate School of Medicine The University of Tokyo  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7195-6905","authenticated-orcid":false,"given":"Gideon Ibrahim","family":"Joseph","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, School of Life Sciences Federal University of Technology  Minna Niger Nigeria"},{"name":"Africa Centre of Excellence for Mycotoxin and Food Safety Federal University of Technology  Minna Niger Nigeria"},{"name":"Department of Parasitology and Virology, Graduate School of Medicine Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edwige Flore","family":"Gouegni","sequence":"additional","affiliation":[{"name":"Africa Centre of Excellence for Mycotoxin and Food Safety Federal University of Technology  Minna Niger Nigeria"},{"name":"College of Pharmacy and Pharmaceutical Sciences Institute of Public Health  Tallahassee Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7121-0713","authenticated-orcid":false,"given":"Israel Ogwuche","family":"Ogra","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Kaduna Nigeria"},{"name":"Biological Institute Tomsk State University  Tomsk Russian Federation"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umar","family":"Saidu","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Kaduna Nigeria"},{"name":"Africa Center of Excellence for Neglected Tropical Diseases and Forensic Biotechnology (ACENTDFB) Ahmadu Bello University  Zaria Kaduna Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oluyinka","family":"Oluseyi Okubanjo","sequence":"additional","affiliation":[{"name":"Department of Veterinary Parasitology and Entomology Ahmadu Bello University  Zaria Kaduna Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adnan Luthfi","family":"Agusta","sequence":"additional","affiliation":[{"name":"Department of Biomedical Chemistry, Graduate School of Medicine The University of Tokyo  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ghulam","family":"Jeelani","sequence":"additional","affiliation":[{"name":"Department of Biomedical Chemistry, Graduate School of Medicine The University of Tokyo  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0680-3889","authenticated-orcid":false,"given":"Mohammed Auwal","family":"Ibrahim","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Kaduna Nigeria"},{"name":"Africa Center of Excellence for Neglected Tropical Diseases and Forensic Biotechnology (ACENTDFB) Ahmadu Bello University  Zaria Kaduna Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,2,23]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.2478\/s11686\u2010020\u201000211\u20104"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid3001.231319"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2006.10.002"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10811\u2010023\u201003181\u2010y"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1177\/004947559402400204"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2013.10.013"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13071\u2010020\u20103984\u20108"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.3791\/58415"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13071\u2010022\u201005352\u20101"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0012111"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0009929"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2025.107526"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20104894(76)90110\u20107"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2023.107138"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11686\u2010021\u201000442\u2010z"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20104894(70)90120\u20107"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007051"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2023.115226"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2017.05.018"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1177\/11786302241274698"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/pim.70033"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70327","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70327","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70327","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T14:43:51Z","timestamp":1775659431000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70327"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2]]},"references-count":21,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2026,2]]}},"alternative-id":["10.1002\/cpz1.70327"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70327","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]]},"article-number":"e70327"}}