title: Report from the HLA class II typing by PCR-SSP Multicentre Study.
abstract: Results from 360 HLA-DR and -DQ 'low-resolution' typings with polymerase chain reaction sequence-specific primers (PCR-SSP), performed by nine laboratories, were analysed for their overall utility in routinely defining the HLA-DR1-DR18, DR51-DR53 and DQ1-DQ9 specificities in less than 2.5 h. Thirty EDTA blood samples and 10 DNA samples were distributed and analysed by each laboratory. DNA was extracted using a rapid bromide salt extraction protocol. Complete HLA-DR and -DQ typings were performed, three by three, on pre-aliquoted 96-tube PCR trays. When compared with reference typing, 351/360 (98%) correct DR typings were obtained, whereas 320/360 (89%) of the DQ phenotypes were correctly assigned. The time for three complete HLA-DR and -DQ 'low-resolution' typings, including DNA extraction, ranged from 2.0 h to 2.3 h. Unfortunately, an unusually high level of PCR amplification failures was observed (3%), probably due to diffusion and a significant volume loss from some of the pre-aliquoted primer mixes. Consequently, only 52% of the typings were without any amplification failure, and 0-2 amplification failures where found in 88% of the PCR-SSP typings performed. The number of HLA-DR-DQ retypings needed was 7 and 8%, respectively, reflecting the low number of typings where allelic identification was directly affected by the relatively high level of amplification failures in this study. Thus, a 91-98% success rate of correctly identified HLA-DR and -DQ alleles could be maintained, even under suboptimal typing conditions.
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: H
authors.initials: H
authors.lastname: Zetterquist
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Bengtsson
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: G
authors.initials: G
authors.lastname: Bäckström
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: I
authors.initials: I
authors.lastname: Egle-Jansson
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: A-M
authors.initials: AM
authors.lastname: Ekdahl
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: N
authors.initials: N
authors.lastname: Grunnet
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: I
authors.initials: I
authors.lastname: Gustafsson
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: I
authors.initials: I
authors.lastname: Knutsen
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Kuhle
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: L
authors.initials: L
authors.lastname: Rydberg
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Spurkland
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: R
authors.initials: R
authors.lastname: Steffensen
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Storgärds
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: E
authors.initials: E
authors.lastname: Szojmer
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: G
authors.initials: G
authors.lastname: Söderholm
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: B
authors.initials: B
authors.lastname: Thuresson
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: H
authors.initials: H
authors.lastname: Turesson
authors.affiliation: Centre for BioTechnology, Karolinska Institute, NOVUM, Huddinge and Department of Clinical Immunology, Karolinska Institute at Huddinge Hospital, Huddinge, Sweden; ,Department of Clinical Immunology, Academic Hospital, Uppsala, Sweden; ,Department of Forensic Medicine, University Hospital, Linköping, Sweden; ,Regional Centre for Blood Transfusion and Clinical Immunology, Aalborg Hospital, Aalborg, Denmark; ,Regional Blood Centre, Sahlgren's Hospital, Gothenburg, Sweden; ,Institute of Transplantation Immunology, National Hospital, Oslo, Norway; ,Department of Clinical Immunology, University Hospital of Aarhus, Aarhus, Denmark; ,Department of Clinical Immunology, National Hospital, Copenhagen, Denmark; and ,Regional Blood Centre, University Hospital, Lund, Sweden.
authors.forename: O
authors.initials: O
authors.lastname: Olerup
mesh_terms: D000483:Alleles; D004247:DNA / Q000302:isolation & purification; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D005802:Genes, MHC Class II / Q000235:genetics*; D006683:HLA-DQ Antigens / Q000032:analysis* / Q000235:genetics; D006684:HLA-DR Antigens / Q000032:analysis* / Q000235:genetics; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D013048:Specimen Handling
chemical_list: D017931:DNA Primers; D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D004247:DNA