HLA-DP typing by DNA amplification and hybridization with specific oligonucleotides

Giovanna Angelini, Teodorica L. Bugawan, Laura Delfino, Henry A. Erlich, Giovanni Battista Ferrara

Research output: Contribution to journalArticle

Abstract

HLA-DP genotyping was performed using dot-blot analysis with synthetic oligonucleotide probes. Fourteen probes were designed based on the known sequence variations in the polymorphic segments of the DPβ second exon. Each probe was tested against genomic DNA amplified by the polymerase chain reaction, using DPβ-specific primers. A total of 45 HLA homozygous B-cell lines, selected from the Tenth International Histocompatibility Workshop and pretyped for the known DPw specificities, were analyzed. Different hybridization patterns were found for each DPw specificity. The oligonucleotide hybridization performed on DPw-negative B-cell lines gave a pattern distinct from those of known DPw specificities, indicating the presence of novel DP allelic sequences. The use of sequence-specific oligonucleotides combined with DNA amplification allows a simple and reliable genotyping of DP antigens.

Original languageEnglish
Pages (from-to)169-177
Number of pages9
JournalHuman Immunology
Volume26
Issue number3
DOIs
Publication statusPublished - 1989

Keywords

  • aa amino acids
  • HLA human leukocyte antigens
  • PCR polymerase chain reaction
  • PLT primed lymphocyte typing
  • RFLP restriction fragment length polymorphism
  • SDS sodium dodecyl sulfate
  • SSPE standard sodium phosphate EDTA

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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    Angelini, G., Bugawan, T. L., Delfino, L., Erlich, H. A., & Ferrara, G. B. (1989). HLA-DP typing by DNA amplification and hybridization with specific oligonucleotides. Human Immunology, 26(3), 169-177. https://doi.org/10.1016/0198-8859(89)90036-0