{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T01:22:10Z","timestamp":1787016130425,"version":"build-2736575974"},"reference-count":37,"publisher":"Wiley","license":[{"start":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T00:00:00Z","timestamp":1766707200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Despite advances in blood safety, emerging infectious agents continue to pose risks to the blood supply. Traditional nucleic acid testing assays primarily target known pathogens, limiting the detection of novel microbes. Nanopore metagenomic sequencing enables agnostic identification of diverse pathogens, potentially enhancing transfusion safety surveillance.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Study Design and Methods<\/jats:title>\n                    <jats:p>We assessed the analytical performance and limit of detection (LoD) of a nanopore MinION metagenomic sequencing workflow for pathogen detection in clinical plasma samples. DNA\/RNA was extracted from 14 archived samples from individuals with confirmed infections (HIV, HBV, HCV, WNV), followed by cDNA synthesis, barcoded library preparation, and nanopore sequencing. Quantitative reference panels were tested, and data were analyzed using a comprehensive bioinformatics pipeline.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Metagenomic sequencing generated over 2 million reads, with 2.3% of reads mapping to microbial sequences. Key transfusion\u2010transmissible viruses, including West Nile virus (WNV), HIV, and hepatitis C virus (HCV), were reliably detected, while protocol modifications enabled HBV identification. Strain\u2010level characterization identified HIV\u20101 subtype B, HCV genotypes 1a, 2b, 4a, HBV genotype C, and WNV lineage 1A. High viral loads produced extensive genome coverage, while lower loads yielded limited recovery. Co\u2010infections, including human pegivirus (HPgV\u20102) and torque teno virus (TTV), were identified. The workflow detected viral targets at 10\n                      <jats:sup>3<\/jats:sup>\n                      genome copy equivalents (GCEs)\/mL, with &gt;\u200950% genome coverage achieved at 10\n                      <jats:sup>4<\/jats:sup>\n                      GCE\/mL.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>Nanopore metagenomic sequencing enables comprehensive detection of blood\u2010borne pathogens in plasma samples. This approach offers a promising complementary strategy for enhancing transfusion safety surveillance, and the demonstrated strain\u2010level characterization supports its potential utility for blood safety applications.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/trf.70051","type":"journal-article","created":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T13:27:46Z","timestamp":1766755666000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Unbiased metagenomic exploration of transfusion\u2010transmitted infections with nanopore sequencing"],"prefix":"10.1111","author":[{"given":"Viswanath","family":"Ragupathy","sequence":"first","affiliation":[{"name":"Office of Blood Research and Review Center for Biologics Evaluation and Research, Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2188-5166","authenticated-orcid":false,"given":"Kylan","family":"Kelley","sequence":"additional","affiliation":[{"name":"Office of Blood Research and Review Center for Biologics Evaluation and Research, Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiangqin","family":"Zhao","sequence":"additional","affiliation":[{"name":"Office of Blood Research and Review Center for Biologics Evaluation and Research, Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilya","family":"Mazo","sequence":"additional","affiliation":[{"name":"Office of Biostatistics and Pharmocovigilance Center for Biologics Evaluation and Research, Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Indira","family":"Hewlett","sequence":"additional","affiliation":[{"name":"Office of Blood Research and Review Center for Biologics Evaluation and Research, Food and Drug Administration  Silver Spring Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,12,26]]},"reference":[{"key":"e_1_2_10_2_1","unstructured":"Blood Safety and availability (WHO Fact Sheet).https:\/\/www.who.int\/news\u2010room\/fact\u2010sheets\/detail\/blood\u2010safety\u2010and\u2010availabilityAccessed 30 May 2025"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/voxs.12070"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)61809-7"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1423-0410.2010.01426.x"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2009.03035.x"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00022-10"},{"key":"e_1_2_10_8_1","first-page":"1","article-title":"Transfusion\u2010transmitted infections: risks and mitigation strategies for Oropouche virus and other emerging arboviruses in Latin America and the Caribbean","volume":"46","author":"Moreira\u2010Soto A","year":"2025","journal-title":"Lancet reg Health Am"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/vox.12925"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/tme.12724"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(14)61034-5"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid2902.221468"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1537-2995.2007.01248.x"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/php.12311"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens12070911"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-pathmechdis-012418-012751"},{"key":"e_1_2_10_17_1","doi-asserted-by":"crossref","first-page":"15809","DOI":"10.1038\/s41598-020-72808-8","article-title":"Metagenomic analysis and identification of emerging pathogens in blood from healthy donors","volume":"10","author":"Xu M","year":"2020","journal-title":"Sci Rep"},{"key":"e_1_2_10_18_1","first-page":"93","article-title":"Viral metagenomics in blood donations with post\u2010donation illness reports from Brazil","volume":"19","author":"Dos Santos Bezerra R","year":"2021","journal-title":"Blood Transfus"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.onehlt.2023.100602"},{"key":"e_1_2_10_20_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13073-015-0220-9","article-title":"Rapid metagenomic identification of viral pathogens in clinical samples by real\u2010time nanopore sequencing analysis","volume":"7","author":"Greninger AL","year":"2015","journal-title":"Genome Med"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01318-07"},{"key":"e_1_2_10_22_1","unstructured":"Oxford Nanopore Technologies\u2010Ligation Sequencing Protocol.https:\/\/nanoporetech.com\/document\/ligation\u2010sequencing\u2010amplicons\u2010native\u2010barcoding\u2010v14\u2010sqk\u2010nbd114\u201024Accessed 12 Nov 2025."},{"issue":"23","key":"e_1_2_10_23_1","article-title":"Brief protocol for EDGE bioinformatics: analyzing microbial and metagenomic NGS data","volume":"7","author":"Philipson CDK","year":"2017","journal-title":"BioProtoc"},{"key":"e_1_2_10_24_1","unstructured":"RIVM Flavivirus Typing Tool.https:\/\/mpf.rivm.nl\/mpf\/typingtool\/flavivirus\/Accessed 12 Nov 2025."},{"key":"e_1_2_10_25_1","unstructured":"Genome Detective's Virus Typing Tool.https:\/\/www.genomedetective.com\/app\/typingtool\/virus\/Accessed 12 Nov 2025."},{"key":"e_1_2_10_26_1","unstructured":"GraphPad Software Inchttps:\/\/www.graphpad.com\/Accessed 12 Nov 2025."},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature16996"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2017.08.019"},{"key":"e_1_2_10_29_1","first-page":"1","article-title":"Discovering disease\u2010causing pathogens in resource\u2010scarce Southeast Asia using a global metagenomic pathogen monitoring system","volume":"119","author":"Bohl JA","year":"2022","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_2_10_30_1","first-page":"400","article-title":"Viral metagenomics applied to blood donors and recipients at high risk for blood\u2010borne infections","volume":"14","author":"Sauvage V","year":"2016","journal-title":"Blood Transfus"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1007644"},{"key":"e_1_2_10_32_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fcimb.2022.943859","article-title":"Nanopore\u2010based metagenomic sequencing for the rapid and precise detection of pathogens among immunocompromised cancer patients with suspected infections","volume":"12","author":"Deng Q","year":"2022","journal-title":"Front Cell Infect Microbiol"},{"key":"e_1_2_10_33_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/JCM.00963-19","article-title":"Metagenomic nanopore sequencing of influenza virus direct from clinical respiratory samples","volume":"58","author":"Lewandowski K","year":"2019","journal-title":"J Clin Microbiol"},{"key":"e_1_2_10_34_1","first-page":"1","article-title":"Molecular rapid diagnostic testing for bloodstream infections: nanopore targeted sequencing with pathogen\u2010specific primers","volume":"88","author":"Han DS","year":"2024","journal-title":"J Infect"},{"key":"e_1_2_10_35_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jviromet.2023.114784","article-title":"Rapid and sensitive single\u2010sample viral metagenomics using nanopore Flongle sequencing","volume":"320","author":"Pichler I","year":"2023","journal-title":"J Virol Methods"},{"key":"e_1_2_10_36_1","first-page":"1","article-title":"A dual\u2010process of targeted and unbiased nanopore sequencing enables accurate and rapid diagnosis of lower respiratory infections","volume":"98","author":"Guo Y","year":"2023","journal-title":"EBioMedicine"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.02225"},{"key":"e_1_2_10_38_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pgph.0003762","article-title":"A consensus statement on dual purpose pathogen surveillance systems: the always on approach","volume":"4","author":"Westhuizen HM","year":"2024","journal-title":"PLoS Glob Public Health"}],"container-title":["Transfusion"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/trf.70051","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T13:27:49Z","timestamp":1766755669000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/trf.70051"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,26]]},"references-count":37,"alternative-id":["10.1111\/trf.70051"],"URL":"https:\/\/doi.org\/10.1111\/trf.70051","archive":["Portico"],"relation":{},"ISSN":["0041-1132","1537-2995"],"issn-type":[{"value":"0041-1132","type":"print"},{"value":"1537-2995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,26]]},"assertion":[{"value":"2025-07-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-26","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"trf.70051"}}