Quality control project of NGS HLA genotyping for the 17th International HLA and Immunogenetics Workshop |
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| Authors: | Kazutoyo Osoegawa Tamara A. Vayntrub Sabine Wenda Dianne De Santis Konstantinos Barsakis Milena Ivanova Susan Hsu Jonathan Barone Rhonda Holdsworth Mary Diviney Medhat Askar Amanda Willis Dawn Railton Sophie Laflin Ketevan Gendzekhadze Arisa Oki Nicoletta Sacchi Michela Mazzocco Marcelo A. Fernandez-Viña |
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| Affiliation: | 1. Histocompatibility, Immunogenetics, and Disease Profiling Laboratory, Stanford Blood Center, Palo Alto, CA, USA;2. Department of Blood Group Serology and Transfusion Medicine, Medical University of Vienna, Vienna, Austria;3. PathWest, Fiona Stanley Hospital, Murdoch, WA, Australia;4. University of Crete, Biology Department, Heraklion, Greece;5. Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA;6. Alexandrovska Hospital, Sofia, Bulgaria;7. Histocompatibility/Molecular Genetics, American Red Cross, Philadelphia, PA, USA;8. Australian Red Cross Blood Services, Melbourne, Australia;9. Baylor University Medical Center, Dallas, TX, USA;10. Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK;11. City of Hope National Medical Center, Duarte, CA, USA;12. Ente Ospedaliero Ospedali Galliera, Genova, Italy;13. Fondazione I.M.E. Istituto Mediterraneo Di Ematologia, Rome, Italy;14. Health Sciences Center, Kuwait University, Jabriya, Kuwait;15. Hellenic Cord Blood Bank, Athens, Greece;p. Hospital Albert Einstein, Sao Paulo, Brazil;q. Centro de Diagonóstico Biomédico, Hospital Clínic de Barcelona, Barcelona, Spain;r. Illumina, Inc., San Diego, CA, USA;s. Johns Hopkins University School of Medicine, Baltimore, MD, USA;t. Kashi Clinical Laboratories, Inc., Portland, OR, USA;u. McGill University Health Centre, Montreal, QC, Canada;v. New Zealand Blood Service, Epsom, Auckland, New Zealand;w. National H&I Service Development Laboratory NHS Blood and Transplant, London, UK;x. One Lambda, Thermo Fisher Scientific, Canoga Park, CA, USA;y. Palacky University, Faculty of Medicine and Dentistry, Olomouc, Czech Republic;z. Primer Centro Argentino de Immunogenetica (PRICAI), Fundación Favaloro, CABA, Argentina;11. Rogachev Federal Research Centre of Pediatric Hematology,Oncology and Immunology, Moscow, Russian Federation;12. The University of Chicago Medicine, Chicago, IL, USA;13. Tokai University School of Medicine, Kanagawa, Japan;14. University Medical Center Utrecht, Netherlands;15. University of California, Los Angeles, Immunogenetics Center, Los Angeles, CA, USA;16. University of California San Diego, La Jolla, CA, USA;17. University of Miami Miller School of Medicine, USA;18. University of Tokyo, Tokyo, Japan;19. Walter Reed Army Institute of Research, Silver Spring, MD, USA;110. Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, USA;111. All India Institute of Medical Sciences, New Delhi, India;112. Anthony Nolan Research Institute and UCL Cancer Institute, Royal Free Campus, London, UK;113. Bo Fu Rui (BFR) Transplant Diagnostics, Beijing, China;114. GenDx, Utrecht, Netherlands;115. Georgetown University Medical Center, Washington DC, USA;1p. Histocompatibility and Immunogenetics Laboratory, Nantes, France;1q. Transplantation and Immunology, Universitat Tuebingen, Germany;1r. Transplantation Immunology, Ulm, Germany;1s. Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill School of Medicine, NC, USA;1t. Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden;1u. Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, WA, USA;1v. Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA;1w. Stanford Genome Technology Center, Palo Alto, CA, USA;1x. Center for Genetics, Children’s Hospital Oakland Research Institute, Oakland, CA, USA;1y. Department of Human Genetics, McGill University, Montreal, Quebec, Canada |
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| Abstract: | ![]()
The 17th International HLA and Immunogenetics Workshop (IHIW) organizers conducted a Pilot Study (PS) in which 13 laboratories (15 groups) participated to assess the performance of the various sequencing library preparation protocols, NGS platforms and software in use prior to the workshop. The organizers sent 50 cell lines to each of the 15 groups, scored the 15 independently generated sets of NGS HLA genotyping data, and generated “consensus” HLA genotypes for each of the 50 cell lines. Proficiency Testing (PT) was subsequently organized using four sets of 24 cell lines, selected from 48 of 50 PS cell lines, to validate the quality of NGS HLA typing data from the 34 participating IHIW laboratories. Completion of the PT program with a minimum score of 95% concordance at the HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 loci satisfied the requirements to submit NGS HLA typing data for the 17th IHIW projects. Together, these PS and PT efforts constituted the 17th IHIW Quality Control project. Overall PT concordance rates for HLA-A, HLA-B, HLA-C, HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DQB1, HLA-DRB1, HLA-DRB3, HLA-DRB4 and HLA-DRB5 were 98.1%, 97.0% and 98.1%, 99.0%, 98.6%, 98.8%, 97.6%, 96.0%, 99.1%, 90.0% and 91.7%, respectively. Across all loci, the majority of the discordance was due to allele dropout. The high cost of NGS HLA genotyping per experiment likely prevented the retyping of initially failed HLA loci. Despite the high HLA genotype concordance rates of the software, there remains room for improvement in the assembly of more accurate consensus DNA sequences by NGS HLA genotyping software. |
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| Keywords: | Corresponding author at: Histocompatibility, Immunogenetics & Disease Profiling Laboratory, Stanford Blood Center, 3155 Porter Drive, Palo Alto, CA 94304 USA. NGS HLA typing Proficiency testing Reference cell panel Quality control CWD Common and Well Documented FHCRC Fred Hutchinson Cancer Research Center FTP File Transfer Protocol GL Genotype List IHIW International HLA and Immunogenetics Workshop IHWG International Histocompatibility Working Group NGS Next Generation Sequencing PS Pilot Study PT Proficiency Testing QC Quality Control RP Reference Panel |
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