{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T14:51:19Z","timestamp":1791211879071,"version":"4.1.0"},"reference-count":33,"publisher":"Oxford University Press (OUP)","issue":"10","license":[{"start":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100021171","name":"Guangdong Basic and Applied Basic Research Foundation","doi-asserted-by":"publisher","award":["2023A1515110676"],"award-info":[{"award-number":["2023A1515110676"]}],"id":[{"id":"10.13039\/501100021171","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Science and Technology Program of Guangzhou","award":["2024B03J1036"],"award-info":[{"award-number":["2024B03J1036"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Third-generation sequencing (TGS) enables comprehensive detection of rare hemoglobin (Hb) variants due to its long-read capability. However, library preparation is often labor-intensive and time-consuming. This study developed a novel one-step library preparation (OSLP) method for TGS. By integrating barcode sequences into target-specific primers, the approach enables immediate barcoding on amplification, streamlining identification of hemoglobinopathy variants.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A retrospective set of DNA samples (n = 455; 394 adult, 61 prenatal) underwent OSLP targeting of the HBA2\/1, HBG2\/1, HBD, and HBB genes on the Qitan Nanopore platform for variant and structural variation detection. Samples were divided into validation and application cohorts based on prior testing with multiplex ligation-dependent probe amplification (MLPA) and Sanger sequencing alongside Gap-polymerase chain reaction (GAP-PCR)\/reverse dot blot (RDB).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>In the validation cohort, OSLP-TGS showed 100% concordance with routine methods (MLPA, Sanger, GAP-PCR, RDB), including 294 samples for \u03b1-globin genotyping [8 structural variants (SVs), 17 single nucleotide variants (SNVs)\/insertions\u2013deletions (indels)], 138 for \u03b2-globin genotyping (4 SVs, 22 SNVs\/indels), and 61 prenatal samples for combined genotyping. The method directly spanned breakpoints and determined cis\/trans configurations. In the application cohort, OSLP-TGS-Nanopore identified additional variants not covered by routine genotyping: 9 extra \u03b1-globin SNVs in 16\/100 samples and 8 extra \u03b2-globin SNVs in 33\/256 samples. Notably, 3 samples with the \u2013SEA deletion harbored co-inherited novel variants, explaining their disproportionately low Hb A2 levels.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The OSLP-TGS-Nanopore assay provides an efficient, comprehensive solution for detecting rare hemoglobinopathy variants, with a framework extendable to population screening for diverse genetic disorders.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/clinchem\/hvag072","type":"journal-article","created":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T11:33:46Z","timestamp":1780400026000},"page":"1079-1092","source":"Crossref","is-referenced-by-count":0,"title":["Rapid Detection of Hemoglobinopathy Variants Using One-Step Library Preparation and Nanopore Sequencing"],"prefix":"10.1093","volume":"72","author":[{"given":"Xu","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University , Guangzhou ,","place":["China"]},{"name":"Guangzhou Darui Biotechnology Co., Ltd , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shang","family":"Yi","sequence":"additional","affiliation":[{"name":"Department of Guangxi Clinical Medical Research Center for Birth Defects, Guangxi Key Laboratory of Reproductive Health and Birth Defects Prevention and Control, Guangxi Zhuang Autonomous Region Women and Children Care Hospital , Nanning ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingyu","family":"Wei","sequence":"additional","affiliation":[{"name":"Fetal Medicine and Prenatal Diagnosis Department, Zhujiang Hospital, Southern Medical University , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guiyun","family":"Zheng","sequence":"additional","affiliation":[{"name":"Fetal Medicine and Prenatal Diagnosis Department, Zhujiang Hospital, Southern Medical University , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Long","family":"Wu","sequence":"additional","affiliation":[{"name":"Guangzhou Darui Biotechnology Co., Ltd , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangzhou Darui Biotechnology Co., Ltd , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guojun","family":"Ouyang","sequence":"additional","affiliation":[{"name":"Guangzhou Darui Biotechnology Co., Ltd , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"He","sequence":"additional","affiliation":[{"name":"Department of Guangxi Clinical Medical Research Center for Birth Defects, Guangxi Key Laboratory of Reproductive Health and Birth Defects Prevention and Control, Guangxi Zhuang Autonomous Region Women and Children Care Hospital , Nanning ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuexi","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Yang","sequence":"additional","affiliation":[{"name":"Fetal Medicine and Prenatal Diagnosis Department, Zhujiang Hospital, Southern Medical University , Guangzhou ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,6,23]]},"reference":[{"key":"2026100509522257300_hvag072-B1","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1056\/NEJMra2021838","article-title":"\u03b2-Thalassemias","volume":"384","author":"Taher","year":"2021","journal-title":"N Engl J Med"},{"key":"2026100509522257300_hvag072-B2","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1093\/clinchem\/hvab049","article-title":"Prenatal hemoglobinopathy evaluation","volume":"67","author":"Manthei","year":"2021","journal-title":"Clin Chem"},{"key":"2026100509522257300_hvag072-B3","first-page":"704","article-title":"Inherited haemoglobin disorders: an increasing global health problem","volume":"79","author":"Weatherall","year":"2001","journal-title":"Bull World Health Organ"},{"key":"2026100509522257300_hvag072-B4","doi-asserted-by":"crossref","first-page":"2310","DOI":"10.1016\/S0140-6736(22)00536-0","article-title":"Thalassaemia","volume":"399","author":"Kattamis","year":"2022","journal-title":"Lancet"},{"key":"2026100509522257300_hvag072-B5","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/S0140-6736(17)31822-6","article-title":"Thalassaemia","volume":"391","author":"Taher","year":"2018","journal-title":"Lancet"},{"key":"2026100509522257300_hvag072-B6","doi-asserted-by":"crossref","first-page":"101074","DOI":"10.1016\/j.blre.2023.101074","article-title":"Thalassaemia in China","volume":"60","author":"Wang","year":"2023","journal-title":"Blood Rev"},{"key":"2026100509522257300_hvag072-B7","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.hoc.2022.12.001","article-title":"Molecular basis and genetic modifiers of thalassemia","volume":"37","author":"Tesio","year":"2023","journal-title":"Hematol Oncol Clin North Am"},{"key":"2026100509522257300_hvag072-B8","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0103020","article-title":"IthaGenes: an interactive database for haemoglobin variations and epidemiology","volume":"9","author":"Kountouris","year":"2014","journal-title":"PLoS One"},{"key":"2026100509522257300_hvag072-B9","doi-asserted-by":"crossref","first-page":"D1063","DOI":"10.1093\/nar\/gkt911","article-title":"Updates of the HbVar database of human hemoglobin variants and thalassemia mutations","volume":"42","author":"Giardine","year":"2014","journal-title":"Nucleic Acids Res"},{"key":"2026100509522257300_hvag072-B10","doi-asserted-by":"crossref","DOI":"10.1093\/database\/baae080","article-title":"Global Globin Network and adopting genomic variant database requirements for thalassemia","volume":"2024","author":"Halim-Fikri","year":"2024","journal-title":"Database (Oxford)"},{"key":"2026100509522257300_hvag072-B11","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/ejhg.2014.131","article-title":"EMQN Best Practice Guidelines for molecular and haematology methods for carrier identification and prenatal diagnosis of the haemoglobinopathies","volume":"23","author":"Traeger-Synodinos","year":"2015","journal-title":"Eur J Hum Genet"},{"key":"2026100509522257300_hvag072-B12","doi-asserted-by":"crossref","first-page":"S39","DOI":"10.5045\/br.2021.2020332","article-title":"Molecular basis and diagnosis of thalassemia","volume":"56","author":"Lee","year":"2021","journal-title":"Blood Res"},{"key":"2026100509522257300_hvag072-B13","doi-asserted-by":"crossref","first-page":"1908","DOI":"10.1056\/NEJMra1404415","article-title":"The \u03b1-thalassemias","volume":"371","author":"Piel","year":"2014","journal-title":"N Engl J Med"},{"key":"2026100509522257300_hvag072-B14","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1186\/s40246-023-00520-5","article-title":"Next-generation sequencing analysis of the molecular spectrum of thalassemia in Southern Jiangxi, China","volume":"17","author":"Yang","year":"2023","journal-title":"Hum Genomics"},{"key":"2026100509522257300_hvag072-B15","article-title":"Next-generation sequencing improves molecular epidemiological characterization of thalassemia in Chenzhou Region, P.R","volume":"33","author":"Zhang","year":"2019","journal-title":"China. J Clin Lab Anal"},{"key":"2026100509522257300_hvag072-B16","doi-asserted-by":"crossref","first-page":"373","DOI":"10.3390\/diagnostics13030373","article-title":"Next-Generation Sequencing (NGS) and Third-Generation Sequencing (TGS) for the diagnosis of thalassemia","volume":"13","author":"Hassan","year":"2023","journal-title":"Diagnostics (Basel)"},{"key":"2026100509522257300_hvag072-B17","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1038\/s41597-025-04762-x","article-title":"Genomes of Prochlorococcus, synechococcus, bacteria, and viruses recovered from marine picocyanobacteria cultures based on Illumina and Qitan nanopore sequencing","volume":"12","author":"Wu","year":"2025","journal-title":"Sci Data"},{"key":"2026100509522257300_hvag072-B18","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1002\/elps.202300270","article-title":"Nanopore sequencing of forensic short tandem repeats using QNome of Qitan Technology","volume":"45","author":"Yang","year":"2024","journal-title":"Electrophoresis"},{"key":"2026100509522257300_hvag072-B19","doi-asserted-by":"crossref","DOI":"10.46471\/gigabyte.154","article-title":"Efficiently constructing complete genomes with CycloneSEQ to fill gaps in bacterial draft assemblies","volume":"2025","author":"Liang","year":"2025","journal-title":"GigaByte"},{"key":"2026100509522257300_hvag072-B20","doi-asserted-by":"crossref","DOI":"10.1093\/gigascience\/giaf079","article-title":"A near-complete genome assembly of the bearded dragon Pogona vitticeps provides insights into the origin of Pogona sex chromosomes","volume":"14","author":"Guo","year":"2025","journal-title":"Gigascience"},{"key":"2026100509522257300_hvag072-B21","article-title":"Identification of a Novel HLA-G allele, HLA-G*01:62, using PolyseqOne and Oxford Nanopore sequencing","volume":"106","author":"Chen","year":"2025","journal-title":"HLA"},{"key":"2026100509522257300_hvag072-B22","article-title":"Next-generation sequencing-based tools or nanopore-based tools: which is more suitable for short tandem repeats genotyping of nanopore sequencing?","volume":"5","author":"Han","year":"2025","journal-title":"Bioinform Adv"},{"key":"2026100509522257300_hvag072-B23","first-page":"30","article-title":"Third-generation sequencing: the spearhead towards the radical transformation of modern genomics","volume":"12","author":"Athanasopoulou","year":"2021","journal-title":"Life (Basel)"},{"key":"2026100509522257300_hvag072-B24","doi-asserted-by":"crossref","DOI":"10.1016\/j.cca.2023.117624","article-title":"Third-generation sequencing for genetic disease","volume":"551","author":"Ling","year":"2023","journal-title":"Clin Chim Acta"},{"key":"2026100509522257300_hvag072-B25","doi-asserted-by":"crossref","DOI":"10.1016\/j.cca.2023.117619","article-title":"The comprehensive analysis of thalassemia alleles (CATSA) based on single-molecule real-time technology (SMRT) is a more powerful strategy in the diagnosis of thalassemia caused by rare variants","volume":"551","author":"Feng","year":"2023","journal-title":"Clin Chim Acta"},{"key":"2026100509522257300_hvag072-B26","doi-asserted-by":"crossref","DOI":"10.1016\/j.gene.2022.146438","article-title":"Detection of rare thalassemia mutations using long-read single-molecule real-time sequencing","volume":"825","author":"Jiang","year":"2022","journal-title":"Gene"},{"key":"2026100509522257300_hvag072-B27","article-title":"Haplotype-resolved genotyping and association analysis of 1,020 \u03b2-thalassemia patients by targeted long-read sequencing","volume":"12","author":"Ye","year":"2025","journal-title":"Advanced Science"},{"key":"2026100509522257300_hvag072-B28","doi-asserted-by":"crossref","DOI":"10.1016\/j.cca.2024.119749","article-title":"Comprehensive analysis of thalassemia alleles (CATSA) based on third-generation sequencing is a comprehensive and accurate approach for neonatal thalassemia screening","volume":"560","author":"Long","year":"2024","journal-title":"Clin Chim Acta"},{"key":"2026100509522257300_hvag072-B29","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1016\/j.jmoldx.2021.06.008","article-title":"A more universal approach to comprehensive analysis of thalassemia alleles (CATSA)","volume":"23","author":"Liang","year":"2021","journal-title":"J Mol Diagn"},{"key":"2026100509522257300_hvag072-B30","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1093\/clinchem\/hvad073","article-title":"Nanopore third-generation sequencing for comprehensive analysis of hemoglobinopathy variants","volume":"69","author":"Huang","year":"2023","journal-title":"Clin Chem"},{"key":"2026100509522257300_hvag072-B31","doi-asserted-by":"crossref","DOI":"10.1002\/jcla.70044","article-title":"Applying the national genomic DNA reference materials to evaluate the performance of nanopore sequencing in identifying thalassemia variants","volume":"39","author":"Wei","year":"2025","journal-title":"Clinical Laboratory Analysis"},{"key":"2026100509522257300_hvag072-B32","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1093\/clinchem\/hvac180","article-title":"Comparison of single molecule, real-time sequencing and nanopore sequencing for analysis of the size, end-motif, and tissue-of-origin of long cell-free DNA in plasma","volume":"69","author":"Yu","year":"2023","journal-title":"Clin Chem"},{"key":"2026100509522257300_hvag072-B33","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1186\/s12920-024-02014-2","article-title":"Application of third-generation sequencing technology for identifying rare \u03b1- and \u03b2-globin gene variants in a Southeast Chinese region","volume":"17","author":"Zhuang","year":"2024","journal-title":"BMC Med Genomics"}],"container-title":["Clinical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/clinchem\/article-pdf\/72\/10\/1079\/68587130\/hvag072.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/clinchem\/article-pdf\/72\/10\/1079\/68587130\/hvag072.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T13:52:32Z","timestamp":1791208352000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/clinchem\/article\/72\/10\/1079\/8714251"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,23]]},"references-count":33,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,6,23]]},"published-print":{"date-parts":[[2026,10,5]]}},"URL":"https:\/\/doi.org\/10.1093\/clinchem\/hvag072","relation":{},"ISSN":["0009-9147","1530-8561"],"issn-type":[{"value":"0009-9147","type":"print"},{"value":"1530-8561","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,10]]},"published":{"date-parts":[[2026,6,23]]}}}