DNA typing by microbead arrays and PCR-SSP: apparent false-negative or -positive hybridization or amplification signals disclose new HLA-B and -DRB1 alleles

Tissue Antigens. 2008 Mar;71(3):238-41. doi: 10.1111/j.1399-0039.2007.00993.x. Epub 2008 Jan 7.

Abstract

Human leukocyte antigen (HLA) typing by polymerase chain reaction-sequence-specific oligonucleotide (PCR-SSO) hybridization on solid phase (microbead assay) or polymerase chain reaction-sequence-specific primers (PCR-SSP) requires interpretation softwares to detect all possible allele combinations. These programs propose allele calls by taking into account false-positive or false-negative signal(s). The laboratory has the option to validate typing results in the presence of strongly cross-reacting or apparent false-negative signals. Alternatively, these seemingly aberrant signals may disclose novel variants. We report here four new HLA-B (B*5620 and B*5716) and HLA-DRB1 alleles (DRB1*110107 and DRB1*1474) that were detected by apparent false-negative or -positive hybridization or amplification patterns, and ultimately resolved by sequencing. To avoid allele misassignments, a comprehensive evaluation of acquired data as documented in a quality assurance system is therefore required to confirm unambiguous typing interpretation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Base Sequence
  • DNA / genetics*
  • DNA Fingerprinting / methods*
  • DNA Fingerprinting / statistics & numerical data
  • DNA Primers / genetics
  • False Negative Reactions
  • False Positive Reactions
  • HLA-B Antigens / genetics*
  • HLA-DR Antigens / genetics*
  • HLA-DRB1 Chains
  • Histocompatibility Testing / methods*
  • Histocompatibility Testing / statistics & numerical data
  • Humans
  • Microspheres
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction / methods
  • Sequence Homology, Nucleic Acid

Substances

  • DNA Primers
  • HLA-B Antigens
  • HLA-DR Antigens
  • HLA-DRB1 Chains
  • DNA