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Several studies have assessed the utilisation of TGS in HLA genotyping, though many of these studies have described the high error rate as an obstacle to achieving a robust and highly accurate HLA typing assay. In 2021, Oxford Nanopore Technologies (ONT) released the high\u2010accuracy sequencing Kit 14 and the MinION flow cell model R10.4.1, which were reported to achieve sequencing accuracies up to 99%. The aim of this study was to validate this novel high\u2010accuracy sequencing kit for HLA genotyping coupled with a full\u2010gene HLA PCR assay. Comparison with historical data obtained using legacy flow cell models such as R9.4, R10.3 and R10.4 was also done to assess progressive improvement in sequencing performance with each sequential release. The workflow was validated based on data throughput, sequence quality and accuracy, and HLA genotyping resolution. An initial validation was performed using an internal reference panel of 42 samples representing common HLA allele groups, followed by an analysis of data obtained from 111 sequencing batch runs since the implementation, to assess assay performance and define quality control metrics to assess inter\u2010run variability and monitor quality. Furthermore, challenges arising from MinION flow cell stability and use, and assessment of barcode contamination are discussed. The findings of this study highlight advantages of ONT sequencing kit 14\/R10.4.1 for HLA genotyping and the implementation considerations for the routine diagnostic HLA laboratory.<\/jats:p>","DOI":"10.1111\/tan.15725","type":"journal-article","created":{"date-parts":[[2024,10,22]],"date-time":"2024-10-22T12:12:08Z","timestamp":1729599128000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Revolutionising High Resolution <scp>HLA<\/scp> Genotyping for Transplant Assessment: Validation, Implementation and Challenges of Oxford Nanopore Technologies' <scp>Q20<\/scp><sup>+<\/sup> Sequencing"],"prefix":"10.1111","volume":"104","author":[{"given":"Naser","family":"El\u2010Lagta","sequence":"first","affiliation":[{"name":"Department of Clinical Immunology PathWest Laboratory Medicine WA, Fiona Stanley Hospital  Perth Australia"},{"name":"School of Biomedical Sciences the University of Western Australia  Perth 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