Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP)

Hum Immunol. 1993 Aug;37(4):201-6. doi: 10.1016/0198-8859(93)90502-r.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Alleles
  • Base Sequence
  • DNA / analysis
  • DNA Primers / chemistry
  • Electrophoresis, Agar Gel
  • False Positive Reactions
  • Genotype
  • HLA-DQ Antigens / classification
  • HLA-DQ Antigens / genetics*
  • HLA-DQ beta-Chains
  • Histocompatibility Testing
  • Humans
  • Kidney Transplantation
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length
  • Tissue Donors

Substances

  • DNA Primers
  • HLA-DQ Antigens
  • HLA-DQ beta-Chains
  • HLA-DQbeta antigen
  • DNA