Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP).
Molecular genotyping of HLA class II genes using group-specific DNA amplification by the PCR followed by probing with (PCR-SSO) probes is too time consuming for the typing of cadaveric organ donors. Recently, amplification of DNA using PCR-SSP has proved a reliable and rapid method for typing HLA-DRB1 genes. PCR-SSP takes 2 hours to perform and is therefore suitable for the genotyping of cadaveric donors. We have designed a set of primers that in eight PCR reactions will positively identify the HLA-DQB1 alleles corresponding to the serologically defined series HLA-DQ2, DQ4, DQ5, DQ6, DQ7, DQ8, and DQ9. Presently, 30 homozygous cell lines and 138 individuals have been typed by the DQB1 PCR-SSP technique and compared with a combination of serology and RFLP with 100% concordance. No false-negative or false-positive amplifications were recorded. All combinations of DQB1 can be readily identified. DQB1 PCR-SSP can take as little as 130 minutes from start to finish, including DNA preparation.
Bunce
M
M
Nuffield Department of Surgery, Oxford Transplant Centre, Churchill Hospital, England.
Taylor
C J
CJ
Welsh
K I
KI
D000483:Alleles; D001483:Base Sequence; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005189:False Positive Reactions; D005838:Genotype; D006683:HLA-DQ Antigens / Q000145:classification / Q000235:genetics*; D059866:HLA-DQ beta-Chains; D006650:Histocompatibility Testing; D006801:Humans; D016030:Kidney Transplantation; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D014019:Tissue Donors
D017931:DNA Primers; D006683:HLA-DQ Antigens; D059866:HLA-DQ beta-Chains; C057615:HLA-DQbeta antigen; D004247:DNA