A 25% error rate in serologic typing of HLA-B homozygotes

D. F. Lorentzen, K. K. Iwanaga, K. J. Meuer, T. L. Moritz, D. I. Watkins

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

The microlymphocytotoxicity technique has been the accepted method for HLA class I typing since the early 1960s. However, it is often difficult to distinguish two related alleles expressed in an individual due to the cross-reactive nature of the alloantibodies used in this technique. This is especially evident at the HLA-B locus, whose more than 180 alleles fall into only 4 major interrelated cross-reactive antigen groups. To estimate the error rate in serologic typing due to the cross-reactive nature of sera, we used polymerase chain reaction with sequence-specific primers (PCR-SSP) amplification to retype 40 individuals who were previously typed as serologic HLA-B locus homozygotes. PCR-SSP revealed that 10 of these 40 individuals (25%) were actually heterozygous at their HLA-B loci. The HLA-B locus alleles of 9 of these 10 discrepant individuals were further analyzed by denaturing gradient gel electrophoresis followed by direct sequencing. The sequence analysis confirmed that all nine individuals were indeed HLA-B locus heterozygotes. This surprisingly high error rate in serologic definition of HLA-B molecules argues for the use of rapid DNA-based techniques in HLA class I typing, even in the setting of solid organ transplantation.

Original languageEnglish (US)
Pages (from-to)359-365
Number of pages7
JournalTissue Antigens
Volume50
Issue number4
DOIs
StatePublished - Jan 1 1997

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Keywords

  • Cross-reaction
  • DGGE
  • HLA-B
  • Homozygote
  • Microlymphocytotoxicity
  • PCR-SSP
  • Serology

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

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