DNA typing with digoxigenin-11-dideoxyuridinetriphosphate-labelled oligonucleotide probes enables non-radioactive analysis using a dual detection system: application for screening HLA-DQA1 polymorphisms

Int Arch Allergy Immunol. 1993;101(1):7-12. doi: 10.1159/000236492.

Abstract

We describe a standardized, highly sensitive, nonradioactive detection procedure for HLA class-II typing of DQA1 alleles and suballeles which has important and cost-effective application in studying HLA disease associations. The procedure involves polymerase chain reaction-based target DNA sequence amplification and dot blotting followed by stringent hybridization with digoxigenin-11-dideoxyuridine triphosphate 3'-end-labelled allele-specific oligonucleotide probes. The dual detection system described here makes this procedure very versatile.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • Deoxyuracil Nucleotides*
  • Dideoxynucleotides
  • Digoxigenin / analogs & derivatives*
  • HLA-DQ Antigens / genetics*
  • HLA-DQ alpha-Chains
  • Histocompatibility Testing* / methods
  • Humans
  • Molecular Sequence Data
  • Oligonucleotide Probes*
  • Polymorphism, Genetic*
  • Sequence Analysis, DNA* / methods

Substances

  • Deoxyuracil Nucleotides
  • Dideoxynucleotides
  • HLA-DQ Antigens
  • HLA-DQ alpha-Chains
  • HLA-DQA1 antigen
  • Oligonucleotide Probes
  • digoxigenin-11-2',3'-dideoxyuridine 5'-triphosphate
  • Digoxigenin