Report from the HLA class II typing by PCR-SSP Multicentre Study.
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.
Zetterquist
H
H
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.
Bengtsson
M
M
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.
Bäckström
G
G
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.
Egle-Jansson
I
I
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.
Ekdahl
A-M
AM
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.
Grunnet
N
N
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.
Gustafsson
I
I
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.
Knutsen
I
I
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.
Kuhle
A
A
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.
Rydberg
L
L
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.
Spurkland
A
A
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.
Steffensen
R
R
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.
Storgärds
M
M
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.
Szojmer
E
E
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.
Söderholm
G
G
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.
Thuresson
B
B
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.
Turesson
H
H
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.
Olerup
O
O
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.
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
D017931:DNA Primers; D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D004247:DNA