Non-radioisotopic typing of human leukocyte antigen class II genes on microplates

Biotechniques. 1993 Nov;15(5):918-25.

Abstract

We describe a new nonradioisotopic method for HLA class II molecular typing performed in 96-well plates of the same size as those used in enzyme-linked immunosorbent assays (ELISA) and radioimmunoassays (RIA). Biotinylated sequence-specific oligonucleotide (SSO) probes are bound to avidin-coated plates. Digoxigenin-labeled PCR-amplified DNA samples are hybridized, washed and detected with a peroxidase conjugated antibody assay. The method was tested by performing a partial HLA DQA1 and DQB1 typing on 69 randomly selected blood samples. The results are completely concordant with a traditional SSO-PCR typing performed on the same samples. This procedure is simple, fast and could be adapted for performance in semi-automated or automated fashion using equipment already available for ELISA and RIA assays.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Avidin
  • Biotin
  • DNA / analysis
  • Diabetes Mellitus, Type 1 / immunology
  • Digoxigenin
  • Genes, MHC Class II*
  • HLA-DQ Antigens / analysis
  • HLA-DQ Antigens / genetics
  • HLA-DQ alpha-Chains
  • HLA-DQ beta-Chains
  • Histocompatibility Antigens Class II / analysis*
  • Histocompatibility Antigens Class II / genetics
  • Histocompatibility Testing / methods*
  • Humans
  • Immunoenzyme Techniques
  • Nucleic Acid Hybridization / methods*
  • Oligonucleotide Probes
  • Polymerase Chain Reaction

Substances

  • HLA-DQ Antigens
  • HLA-DQ alpha-Chains
  • HLA-DQ beta-Chains
  • HLA-DQA1 antigen
  • HLA-DQB1 antigen
  • Histocompatibility Antigens Class II
  • Oligonucleotide Probes
  • Avidin
  • Biotin
  • DNA
  • Digoxigenin