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

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

Research output: Contribution to journalArticle

23 Citations (Scopus)

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
Pages (from-to)359-365
Number of pages7
JournalTissue Antigens
Volume50
Issue number4
StatePublished - Oct 7 1997
Externally publishedYes

Fingerprint

HLA-B Antigens
Homozygote
Alleles
Isoantibodies
Polymerase Chain Reaction
Denaturing Gradient Gel Electrophoresis
Organ Transplantation
Heterozygote
Sequence Analysis
Antigens
DNA
Serum

Keywords

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

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

Lorentzen, D. F., Iwanaga, K. K., Meuer, K. J., Moritz, T. L., & Watkins, D. (1997). A 25% error rate in serologic typing of HLA-B homozygotes. Tissue Antigens, 50(4), 359-365.

A 25% error rate in serologic typing of HLA-B homozygotes. / Lorentzen, D. F.; Iwanaga, K. K.; Meuer, K. J.; Moritz, T. L.; Watkins, David.

In: Tissue Antigens, Vol. 50, No. 4, 07.10.1997, p. 359-365.

Research output: Contribution to journalArticle

Lorentzen, DF, Iwanaga, KK, Meuer, KJ, Moritz, TL & Watkins, D 1997, 'A 25% error rate in serologic typing of HLA-B homozygotes', Tissue Antigens, vol. 50, no. 4, pp. 359-365.
Lorentzen DF, Iwanaga KK, Meuer KJ, Moritz TL, Watkins D. A 25% error rate in serologic typing of HLA-B homozygotes. Tissue Antigens. 1997 Oct 7;50(4):359-365.
Lorentzen, D. F. ; Iwanaga, K. K. ; Meuer, K. J. ; Moritz, T. L. ; Watkins, David. / A 25% error rate in serologic typing of HLA-B homozygotes. In: Tissue Antigens. 1997 ; Vol. 50, No. 4. pp. 359-365.
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